Enzymatic Modification of Saponins

Abstract

Methods for the enzymatic modification of saponins produced in plant cell culture, products made thereby, uses of said products and also to other associated aspects. The saponins may be obtained from plant cell culture extracts of Quillaja species, such as extracts of Quillaja saponaria Molina.

Description

TECHNICAL FIELD

[0001] The present application generally relates to saponins, in particular methods for the enzymatic modification of saponins produced in in vitro plant cell culture, saponin products made thereby, uses of said saponin products and also to other associated aspects. The present application further relates to the use of glucosidases and rhamnosidases, in particular mutated glucosidases and rhamnosidases, in order to increase the cell culture production of saponins of interest. The saponins may be obtainable from plant cells capable of synthesizing saponins, in particular, plant cells originating from Quillaja species, including from plant cell culture extracts thereof.BACKGROUND

[0002] Saponins are steroid or terpenoid glycosides. They have a broad range of uses from fire extinguisher foams to food additives and immunostimulants (Reichert, 2019).

[0003] Saponins have been of interest as immunostimulants for many decades (see, for example, Hyslop, 1969). Traditionally, saponins have been purified from plants, such as for example Quillaja saponaria Molina trees. Quil A is a saponin fraction derived from an aqueous extract from the bark of Quillaja saponaria (Dalsgaard, 1974). Quil A itself contains a plurality of components with the four most predominant Quil A fractions purified by reverse phase chromatography, namely QS-7, QS-17, QS-18 and QS-21, all having immunostimulatory activity although varying in haemolytic activity and toxicity (Kensil, 1991; Kensil, 1995). The main saponin fraction, QS-18, was found to be highly toxic in mice but saponin fractions QS-7 and QS-21 were far less toxic. QS-21, being more abundant than QS-7, has been the most widely studied saponin adjuvant (Ragupathi, 2011).

[0004] Liquid chromatography / mass spectrometry analysis of Quillaja saponaria bark water / methanol extracts has revealed over 100 saponins, many of which have been assigned structures (Nyberg, 2000; Nyberg, 2003; Kite, 2004).

[0005] Quillaja brasiliensis (A St. -Hil & Tul) Mart. extracts have been described, with the identity of various components therein determined by mass spectrometry. Many saponin components in Quillaja brasiliensis extracts correspond to saponins found in Quillaja saponaria extracts (Wallace, 2017; Wallace, 2019) and Quillaja brasiliensis extracts have also been shown to have immunostimulant effects (Cibulski, 2018; Yendo, 2017).

[0006] The Adjuvant System 01 (AS01) is a liposome-based adjuvant which contains two immunostimulants, 3-O-desacyl-4′-monophosphoryl lipid A (3D-MPL) and QS-21 (Garcon, 2011; Didierlaurent, 2017). 3D-MPL is a non-toxic derivative of the lipopolysaccharide from Salmonella minnesota. AS01 is included in vaccines for malaria (RTS,S—Mosquirix™) and Herpes zoster (HZ / su—Shingrix™), and in multiple candidate vaccines. AS01 injection results in rapid and transient activation of innate immunity in animal models. Neutrophils and monocytes are rapidly recruited to the draining lymph node (dLN) upon immunization. Moreover, AS01 induces recruitment and activation of MHCIIhigh dendritic cells (DC), which are necessary for T cell activation (Didierlaurent, 2014). Some data are also available on the mechanism of action of the components of AS01. 3D-MPL signals via TLR4, stimulating NF-κB transcriptional activity and cytokine production and directly activates antigen-presenting cells (APCs) both in humans and in mice (De Becker, 2000; Ismaili, 2002; Martin, 2003; Mata-Haro, 2007). QS-21 promotes high antigen-specific antibody responses and CD8+ T-cell responses in mice (Kensil, 1998; Newman, 1992; Soltysik, 1995) and antigen-specific antibody responses in humans (Livingston, 1994). Because of its physical properties, it is thought that QS-21 might act as a danger signal in vivo (Lambrecht, 2009; Li, 2008). Although QS-21 has been shown to activate ASC-NLRP3 inflammasome and subsequent IL-1p / IL-18 release (Marty-Roix, 2016), the exact molecular pathways involved in the adjuvant effect of saponins have yet to be clearly defined.

[0007] Extracts of Quillaja saponaria are commercially available, including fractions thereof with differing degrees of purity such as Quil A, Fraction A, Fraction B, Fraction C, QS-7, QS-17, QS-18 and QS-21.

[0008] The enzymatic hydrolysis of Quil-A by Rapidase® Revelation Aroma has been described during the development of a quality control method to provide a degraded reference sample (Lecas, 2021).

[0009] The current source for saponins is mostly dependent on natural resources, and such natural resources may be limiting. As an alternative to the bark of Quillaja saponaria, cell-culture-derived approaches have been developed and are expected to represent a more sustainable source of saponins. For example, WO 94 / 10291 discloses cultured cells of Quillaja saponaria and methods for preparing saponins for use as active substances useful as adjuvants. However, saponins of interest so produced may be in low amounts. Furthermore, separation of certain saponins from other components, particularly other saponin components which may have similar structures, can be burdensome. Consequently, there remains a need for new methods which may improve the yield of saponins of interest produced in plant cell culture and / or facilitate removal of undesired saponin components.

[0010] Modestobacter marinus glucosidase (Uniparc reference UPI000260A2FA, Uniprot reference I4EYD5) is a naturally occurring glucosidase. There remains a need for further glucosidases which may have improved properties.

[0011] Kribbella flavida rhamnosidase (Uniparc reference UPI00019BDB13, Uniprot reference D2PMT) is a naturally occurring rhamnosidase. There remains a need for further rhamnosidases which may have improved properties.SUMMARY OF THE INVENTION

[0012] The present invention provides a method for making a product saponin, said method comprising the steps of:

[0013] (i) providing a plant cell culture extract comprising saponins; and

[0014] (ii) enzymatically converting a starting saponin from the plant cell culture extract to a product saponin.

[0015] Further provided is a method for making a product saponin, said method comprising the steps of:

[0016] (i) culturing plant cells capable of synthesizing saponins under conditions leading to the synthesis of saponins;

[0017] (ii) recovering saponins from the plant cell culture; and

[0018] (iii) enzymatically converting a starting saponin from the recovered saponins to the product saponin.

[0019] Further provided is a method for making a product saponin, said method comprising the steps of:

[0020] (i) culturing plant cells capable of synthesizing saponins under conditions leading to the synthesis of saponins;

[0021] (ii) enzymatically converting a starting saponin from the synthesized saponins to the product saponin; and

[0022] (iii) recovering saponins from the plant cell culture.

[0023] Further provided is a method for increasing the amount of a product saponin obtainable from a plant cell culture, said method comprising the steps of:

[0024] (i) culturing plant cells capable of synthesizing saponins under conditions leading to the synthesis of saponins;

[0025] (ii) recovering saponins from the plant cell culture; and

[0026] (iii) enzymatically converting a starting saponin from the recovered saponins to the product saponin.

[0027] Further provided is a method for increasing the amount of a product saponin obtainable from a plant cell culture, said method comprising the steps of:

[0028] (i) culturing plant cells capable of synthesizing saponins under conditions leading to the synthesis of saponins;

[0029] (ii) enzymatically converting a starting saponin from the synthesized saponins to the product saponin; and

[0030] (iii) recovering saponins from the plant cell culture.

[0031] Further provided is a method for reducing the amount of a starting saponin obtainable from a plant cell culture, said method comprising the following steps of:

[0032] (i) culturing plant cells capable of synthesizing saponins under conditions leading to the synthesis of saponins;

[0033] (ii) recovering saponins from the plant cell culture; and

[0034] (iii) enzymatically converting the starting saponin from the recovered saponins to a product saponin.

[0035] Further provided is a method for reducing the amount of a starting saponin obtainable from a plant cell culture, said method comprising the following steps of:

[0036] (i) culturing plant cells capable of synthesizing saponins under conditions leading to the synthesis of saponins;

[0037] (ii) enzymatically converting a starting saponin from the synthesized saponins to the product saponin; and

[0038] (iii) recovering saponins from the plant cell culture.

[0039] Further provided is a method for producing saponins by plant cell culture, said method comprising the steps of:

[0040] (i) culturing plant cells capable of synthesizing saponins under conditions leading to the synthesis of saponins; and

[0041] (ii) recovering saponins from the plant cell culture, wherein the yield of a product saponin is increased by enzymatically converting a starting saponin from recovered saponins to the product saponin.

[0042] The use of a glycosidase and / or a rhamnosidase for enzymatically converting a starting saponin obtained from a plant cell culture to a product saponin according to the methods of the invention is also provided by the invention.

[0043] Suitably, the glucosidase is an enzyme comprising, such as consisting of: (i) an amino acid sequence according to SEQ ID No. 262, 208, 63, 229, 250, 5, 101, 207, 169, 247, 302, 324, 319, 9,240, 325, 338, 850, 879, 868, 826, 804, 888, 881, 891, 816, 827, 857, 853, 842, 814, 886, 885, 838, 829, 808, 828, 870, 873, 844, 882, 874, 825, 824, 823, 810, 894, 849, 803, 890, 841, 832, 830, 845, 871, 837, 883 or 809 or functional variants thereof; or (ii) an amino acid sequence according to SEQ ID No. 262, 208, 63, 229, 250, 5, 101, 207, 169, 247, 302, 324, 319, 9,240, 325, 338, 850, 879, 868, 826, 804, 888, 881, 891, 816, 827, 857, 853, 842, 814, 886, 885, 838, 829, 808, 828, 870, 873, 844, 882, 874, 825, 824, 823, 810, 894, 849, 803, 890, 841, 832, 830, 845, 871, 837, 883 or 809 or functional variants thereof

[0044] Suitably, the rhamnosidase is an enzyme comprising, such as consisting of, an amino acid sequence according to SEQ ID No. 992, 1003, 1052, 1073, 1017, 1055, 1075, 1001, 1007, 1061, 1079, 1027, 1039, 1041, 989, 1053, 1018, 1066, 1082, 1076, 993, 1077, 1046, 1015, 1063, 1054, 1074, 1067 or 1033, or functional variants thereof.

[0045] The use of an engineered glucosidase and / or an engineered rhamnosidase polypeptides for enzymatically converting a starting saponin obtained from a plant cell culture to a product saponin according to the methods of the invention is also provided by the invention.

[0046] Suitably, the engineered glucosidase polypeptide comprises, such as consists of, an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID No. 262, or a functional fragment thereof, wherein the engineered glucosidase polypeptide includes at least one residue substitution from:

[0047] F44Y;

[0048] V60L;

[0049] G117A;

[0050] F170N;

[0051] V263G or V263L;

[0052] N351H or N351Q;

[0053] A355H, A355I, A355L, A355M, A355R, A355T or A355W;

[0054] A356P;

[0055] R357A, R357C, R357K, R357M or R357Q;

[0056] G362C;

[0057] T365A, T365N or T365S;

[0058] L367C;

[0059] V394R;

[0060] V395Y;

[0061] Q396E, Q396G, Q396N, Q396P, Q396R, Q396S or Q396Y;

[0062] F430W;

[0063] R435F;

[0064] V438T;

[0065] V440F;

[0066] F442M or F442Q;

[0067] G444T;

[0068] A473F or A473R;

[0069] L474C, L474I or L474V;

[0070] I475F;

[0071] L492C, L492G, L492H, L492I, L492N, L492Q, L492V, L492W or L492Y;

[0072] Q493F or Q493H;

[0073] P494H or P494I;

[0074] S495I, S495K or S495Q;

[0075] G496P or G496W;

[0076] D498A, D498E, D498F, D498I, D498K, D498L, D498N, D498P, D498R, D498S, D498T or D498V;

[0077] A502R;

[0078] M504G or M504R;

[0079] L507A or L507R;

[0080] T508M;

[0081] L529M;

[0082] F535P;

[0083] A536D or A536E;

[0084] A537R;

[0085] F541A, F541I, F541L, F541M or F541V;

[0086] L542I;

[0087] Q543G or Q543L;

[0088] E547L; and

[0089] Y585W.

[0090] Suitably, the engineered rhamnosidase polypeptide comprises, such as consists of, an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID No. 1017, or a functional fragment thereof, wherein the engineered rhamnosidase polypeptide includes at least one residue substitution from:

[0091] (i) A56C

[0092] (ii) A143P

[0093] (iii) Q181H, Q181R or Q181S

[0094] (iv) L214M

[0095] (v) G215S

[0096] (vi) F216M

[0097] (vii) G218D or G218N

[0098] (viii) K219G

[0099] (ix) A238M

[0100] (x) T252Y

[0101] (xi) T311W

[0102] (xii) V326C

[0103] (xiii) G357C

[0104] (xiv) S369C, S369I, S369K or S369M

[0105] (xv) I487M, I487Q or I487V

[0106] (xvi) K492N

[0107] (xvii) V499T

[0108] (xviii) G508S

[0109] (xix) R543C

[0110] (xx) L557Y

[0111] (xxi) G634A

[0112] (xxii) S635N

[0113] (xxiii) A690C and

[0114] (xxiv) Q921H.

[0115] The invention also provides a saponin prepared by the methods herein, a saponin-containing composition comprising a product saponin prepared by the methods herein, adjuvant compositions comprising said saponins or saponin-containing compositions, and adjuvant compositions prepared using said saponins or saponin-containing compositions.

[0116] Also provided is a method for preparing an adjuvant comprising the steps of preparing a saponin or a saponin-containing composition from a plant cell culture extract according to the method of the invention, and formulating the saponin or saponin-containing composition into an adjuvant.

[0117] Use of a saponin or saponin-containing composition obtained from a plant cell culture extract of the invention in the manufacture of an adjuvant composition is also provided.

[0118] Further provided are immunogenic compositions comprising a saponin or saponin containing-composition according to the invention and an antigen or a polynucleotide encoding an antigen.

[0119] Also provided is a method for preparing an immunogenic composition comprising the steps of preparing an adjuvant according to the method of the invention, and formulating it into a composition comprising an antigen or a polynucleotide encoding an antigen.

[0120] Kits comprising (i) a saponin or saponin-containing composition according to the invention and (ii) an antigen or a polynucleotide encoding an antigen are also provided.BRIEF DESCRIPTION OF THE FIGURES

[0121] FIG. 1 HPLC chromatogram of a crude aqueous Quillaja saponaria bark extract

[0122] FIG. 2 HPLC-UV chromatogram of a crude aqueous Quillaja saponaria bark extract

[0123] FIG. 3 UPLC-UV chromatogram of a crude aqueous Quillaja saponaria bark extract

[0124] FIG. 4 UPLC-UV chromatogram of a polystyrene purified Quillaja saponaria QS-21 saponin extract with low content of 2018 component

[0125] FIG. 5 UPLC-UV / MS chromatogram of a Quillaja saponaria QS-21 purified saponin extract with low content of 2018 component

[0126] FIG. 6 UPLC-UV / MS chromatogram detail of a Quillaja saponaria QS-21 purified saponin extract with low content of 2018 component

[0127] FIGS. 7A & 7B Extracted mass chromatograms for 1988 (FIG. 7A) and 2002 (FIG. 7B) molecular weight ions of a Quillaja saponaria QS-21 purified saponin extract with low content of 2018 component

[0128] FIG. 8 Combined centroid spectrum of Quillaja saponaria QS-21 purified saponin extract with low content of 2018 component

[0129] FIG. 9 UPLC-UV chromatogram of Quillaja saponaria QS-21 purified saponin extract with low 2018 component

[0130] FIGS. 10A & 10B LCMS / MS chromatogram for QS-18 2150 (FIG. 10A) and QS-21 1988 (FIG. 10B) component content in QS-21 standard in Example 4-2

[0131] FIGS. 11A & 11B LCMS / MS chromatogram for QS-18 2150 (FIG. 11A) and QS-21 1988 (FIG. 11B) component content following negative control treatment in Example 4-2

[0132] FIGS. 12A & 12B LCMS / MS chromatogram for QS-18 2150 (FIG. 12A) and QS-21 1988 (FIG. 12B) component content following glucosidase SEQ ID No. 262 treatment in Example 4-2

[0133] FIG. 13 UV HPLC chromatogram following glucosidase SEQ ID No. 262 treatment (upper trace) and negative control treatment (lower trace) of Crude Bark Extract (CBE) in Example 4-4

[0134] FIGS. 14A & 14B LCMS / MS chromatogram for QS-17 2296 (FIG. 14A) and QS-18 2150 (FIG. 14B) component content following negative control treatment in Example 6-1

[0135] FIGS. 15A & 15B LCMS / MS chromatogram for desglucosyl-QS-17 2134 (FIG. 15A) and QS-21 1988 (FIG. 15B) component content following negative control treatment in Example 6-1

[0136] FIGS. 16A & 16B LCMS / MS chromatogram for QS-17 2310 (FIG. 16A) and QS-18 2164 (FIG. 16B) component content following negative control treatment in Example 6-1

[0137] FIGS. 17A & 17B LCMS / MS chromatogram for QS-17 2296 (FIG. 17A) and QS-18 2150 (FIG. 17B) component content following rhamnosidase SEQ ID No. 1017 treatment in Example 6-1

[0138] FIGS. 18A & 18B LCMS / MS chromatogram for desglucosyl-QS-17 2134 (FIG. 18A) and QS-21 1988 (FIG. 18B) component content following rhamnosidase SEQ ID No. 1017 treatment in Example 6-1

[0139] FIGS. 19A & 19B LCMS / MS chromatogram for QS-17 2310 (FIG. 19A) and QS-18 2164 (FIG. 19B) component content following rhamnosidase SEQ ID No. 1017 treatment in Example 6-1

[0140] FIG. 20 UV HPLC chromatogram following rhamnosidase SEQ ID No. 1017 treatment (upper trace) and negative control treatment (lower trace) of Treated Bark Extract (TBE) in Example 6-2

[0141] FIG. 21 UV HPLC chromatogram following rhamnosidase SEQ ID No. 1017 treatment (upper trace) and negative control treatment (lower trace) of CBE in Example 6-3

[0142] FIGS. 22A & 22B LCMS / MS chromatogram for QS-21 1988 component content at TO (FIG. 22A) and at 24 hrs (FIG. 22B) following dual enzyme treatment of CBE in Example 7

[0143] FIGS. 23A & 23B Illustrative UV HPLC chromatograms following glucosidase enzyme treatment of CBE (FIG. 23A) and negative control treatment of CBE (FIG. 23B) in Example 8

[0144] FIG. 24 Illustrative UV HPLC chromatogram following rhamnosidase enzyme treatment of CBE (upper trace) and negative control treatment of CBE (lower trace) in Example 9

[0145] FIG. 25 HPLC-UV chromatogram of untreated and enzyme treated CBE at 1 L scale from Example 11

[0146] FIG. 26 UPLC-UV chromatogram following purification of untreated and enzyme treated CBE at 1 L scale from Example 11 (full acquisition) FIG. 27 UPLC-UV chromatogram following purification of untreated and enzyme treated CBE at 1 L scale from Example 11 (zoom)

[0147] FIG. 28 Saponin productivity of plant cell cultures untreated and dual enzyme-treated from Example 12. The content of the indicated saponin component was analysed and measured by LCMS / MS in a Crude Cell extract (CCE) obtained from the indicated plant cell culture. The corresponding saponin component concentration (expressed in ug / ml of culture) was retrospectively calculated relative to the indicated plant cell culture. To facilitate visualization, the concentrations of each analysed saponin component have been regrouped within one single bar in the bar graph. The identity of the saponin components analysed is indicated by the colour code provided in the figure.

[0148] FIG. 29 Saponin profile and proportion in a plant cell culture extract. The indicated saponin components have been analysed and measured by LCMS / MS at the indicated steps of the process described in Example 13: (i) pre-enzymation; (ii) post-enzymation; (iii) post-UF / DF concentration; (iv) and post-phenyl chromatography. Results are presented as the respective percentage of a given saponin component, as compared with the sum of the percentages of all saponin components analysed (the total amounting to 100%). The identity of the saponin components analysed is indicated by the colour code provided in the figure.BRIEF DESCRIPTION OF THE SEQUENCE IDENTIFIERSSEQ ID NO: 1: Amino acid sequence for Cyberlindnera fabianii glucosidase, Uniparc reference UPI00049B1A8C, Uniprot reference A0A061B3J2.

[0150] SEQ ID NO: 2: Amino acid sequence for Flavobacterium gilvum glucosidase, Uniparc reference UPI0004E3EF7B, Uniprot reference A0A085EII0.

[0151] SEQ ID NO: 3: Amino acid sequence for Algibacter lectus glucosidase, Uniparc reference UPI00050EE490, Uniprot reference A0A090X649.

[0152] SEQ ID NO: 4: Amino acid sequence for Microbacterium azadirachtae glucosidase, Uniparc reference UPI0005ECB51E, Uniprot reference A0A0F0LB94.

[0153] SEQ ID NO: 5: Amino acid sequence for Actinobacteria bacterium glucosidase, Uniparc reference UPI0006588DAD, Uniprot reference A0A0J0UT37.

[0154] SEQ ID NO: 6: Amino acid sequence for Chloroflexi bacterium glucosidase, Uniparc reference UPI0007968552, Uniprot reference A0A136KWB3.

[0155] SEQ ID NO: 7: Amino acid sequence for Komagataeibacter rhaeticus glucosidase, Uniparc reference UPI0002080410, Uniprot reference A0A181C809.

[0156] SEQ ID NO: 8: Amino acid sequence for Bacteroides sp. glucosidase, Uniparc reference UPI0008211BFC, Uniprot reference A0A1C5WEL8.

[0157] SEQ ID NO: 9: Amino acid sequence for Streptomyces rubrolavendulae glucosidase, Uniparc reference UPI00085A2BD0, Uniprot reference A0A1D8FZW3.

[0158] SEQ ID NO: 10: Amino acid sequence for Clostridium roseum glucosidase, Uniparc reference UPI00098C60F6, Uniprot reference A0A1S8KYM5.

[0159] SEQ ID NO: 11: Amino acid sequence for uncultured bacterium glucosidase, Uniparc reference UPI0009CE0D4C, Uniprot reference A0A1V5M6V6.

[0160] SEQ ID NO: 12: Amino acid sequence for Firmicutes bacterium glucosidase, Uniparc reference UPI0009D4127D, Uniprot reference A0A1V6AN95.

[0161] SEQ ID NO: 13: Amino acid sequence for Anthracocystis flocculosa glucosidase, Uniparc reference UPI00045601AB, Uniprot reference A0A061H1Z3.

[0162] SEQ ID NO: 14: Amino acid sequence for Bifidobacterium boum glucosidase, Uniparc reference UPI0004FF77C7, Uniprot reference A0A086ZKU2.

[0163] SEQ ID NO: 15: Amino acid sequence for Jejuia pallidilutea glucosidase, Uniparc reference UPI00051EDBDE, Uniprot reference A0A098LTR2.

[0164] SEQ ID NO: 16: Amino acid sequence for Ceratocystis fimbriata glucosidase, Uniparc reference UPI00062105AB, Uniprot reference A0A0F8B2B0.

[0165] SEQ ID NO: 17: Amino acid sequence for Actinobacteria bacterium glucosidase, Uniparc reference UPI0006583AB1, Uniprot reference A0A0J0UVW7.

[0166] SEQ ID NO: 18: Amino acid sequence for Rhodococcus sp. glucosidase, Uniparc reference UPI0007AABFAD, Uniprot reference A0A143QAX3.

[0167] SEQ ID NO: 19: Amino acid sequence for Valsa mali glucosidase, Uniparc reference UPI0007F2D02D, Uniprot reference A0A194VF47.

[0168] SEQ ID NO: 20: Amino acid sequence for uncultured Bacteroides sp. glucosidase, Uniparc reference UPI000821004D, Uniprot reference A0A1C5WSI4.

[0169] SEQ ID NO: 21: Amino acid sequence for Eisenbergiella tayi glucosidase, Uniparc reference UPI00084089B2, Uniprot reference A0A1E3ALT2.

[0170] SEQ ID NO: 22: Amino acid sequence for Streptomyces sp. glucosidase, Uniparc reference UPI000978E914, Uniprot reference A0A1V2MYI4.

[0171] SEQ ID NO: 23: Amino acid sequence for Firmicutes bacterium glucosidase, Uniparc reference UPI0009CBF21C, Uniprot reference A0A1V5MH90.

[0172] SEQ ID NO: 24: Amino acid sequence for Tenericutes bacterium glucosidase, Uniparc reference UPI0009D5B1F0, Uniprot reference A0A1V6BAK3.

[0173] SEQ ID NO: 25: Amino acid sequence for Gluconobacter oxydans glucosidase, Uniparc reference UPI0004A87350, Uniprot reference A0A067Z479.

[0174] SEQ ID NO: 26: Amino acid sequence for Bifidobacterium catenulatum glucosidase, Uniparc reference UPI00050787A2, Uniprot reference A0A087B8Q8.

[0175] SEQ ID NO: 27: Amino acid sequence for Bionectria ochroleuca glucosidase, Uniparc reference UPI0005965863, Uniprot reference A0A0B7K538.

[0176] SEQ ID NO: 28: Amino acid sequence for Parcubacteria sp. glucosidase, Uniparc reference UPI0006377CA4, Uniprot reference A0A0G0GD78.

[0177] SEQ ID NO: 29: Amino acid sequence for Microbacterium ketosireducens glucosidase, Uniparc reference UPI0006228575, Uniprot reference A0A0M2H276.

[0178] SEQ ID NO: 30: Amino acid sequence for Roseburia faecis glucosidase, Uniparc reference UPI0006C454B4, Uniprot reference A0A173R3W4.

[0179] SEQ ID NO: 31: Amino acid sequence for Kwoniella dejecticola glucosidase, Uniparc reference UPI0007F1D695, Uniprot reference A0A1A6A050.

[0180] SEQ ID NO: 32: Amino acid sequence for uncultured Clostridium sp. glucosidase, Uniparc reference UPI0008232A70, Uniprot reference A0A1C6C862.

[0181] SEQ ID NO: 33: Amino acid sequence for Cyberlindnera jadinii glucosidase, Uniparc reference UPI000866CB30, Uniprot reference A0A1E4S2F8.

[0182] SEQ ID NO: 34: Amino acid sequence for Bacteroidetes bacterium glucosidase, Uniparc reference UPI0009D3483D, Uniprot reference A0A1V5G4W6.

[0183] SEQ ID NO: 35: Amino acid sequence for Verrucomicrobia bacterium glucosidase, Uniparc reference UPI0009CC9AEA, Uniprot reference A0A1V5Q4R4.

[0184] SEQ ID NO: 36: Amino acid sequence for Bacteroidetes bacterium glucosidase, Uniparc reference UPI0009C6DE73, Uniprot reference A0A1V6BV25.

[0185] SEQ ID NO: 37: Amino acid sequence for Lichtheimia ramosa glucosidase, Uniparc reference UPI0004E051A9, Uniprot reference A0A077WUK7.

[0186] SEQ ID NO: 38: Amino acid sequence for Bifidobacterium mongoliense glucosidase, Uniparc reference UPI0005060F52, Uniprot reference A0A087BWT7.

[0187] SEQ ID NO: 39: Amino acid sequence for Vibrio ishigakensis glucosidase, Uniparc reference UPI000591CED9, Uniprot reference A0A0B8NZY1.

[0188] SEQ ID NO: 40: Amino acid sequence for Phaeomoniella chlamydospora glucosidase, Uniparc reference UPI00063B706F, Uniprot reference A0A0G2HEV5.

[0189] SEQ ID NO: 41: Amino acid sequence for Ardenticatena maritima glucosidase, Uniparc reference UPI0006C04F59, Uniprot reference A0A0M8K5H7.

[0190] SEQ ID NO: 42: Amino acid sequence for Coprococcus comes glucosidase, Uniparc reference UPI000197E031, Uniprot reference A0A173WPC4.

[0191] SEQ ID NO: 43: Amino acid sequence for Nocardioides dokdonensis glucosidase, Uniparc reference UPI0007DDBAB3, Uniprot reference A0A1A9GNJ0.

[0192] SEQ ID NO: 44: Amino acid sequence for uncultured Clostridium sp. glucosidase, Uniparc reference UPI000822F7EB, Uniprot reference A0A1C6EBC1.

[0193] SEQ ID NO: 45: Amino acid sequence for Acetobacterium wieringae glucosidase, Uniparc reference UPI000878EB40, Uniprot reference A0A1F2PFB4.

[0194] SEQ ID NO: 46: Amino acid sequence for Tenericutes bacterium glucosidase, Uniparc reference UPI0009D0A2C1, Uniprot reference A0A1V5HNJ3.

[0195] SEQ ID NO: 47: Amino acid sequence for Tenericutes bacterium glucosidase, Uniparc reference UPI0009CB8A73, Uniprot reference A0A1V5UFI1.

[0196] SEQ ID NO: 48: Amino acid sequence for uncultured bacterium glucosidase, Uniparc reference UPI0009CC90AF, Uniprot reference A0A1V6CDT2.

[0197] SEQ ID NO: 49: Amino acid sequence for Parabacteroides distasonis glucosidase, Uniparc reference UPI0004D8E473, Uniprot reference A0A078SYD0.

[0198] SEQ ID NO: 50: Amino acid sequence for Bifidobacterium psychraerophilum glucosidase, Uniparc reference UPI0005006C6A, Uniprot reference A0A087CJB0.

[0199] SEQ ID NO: 51: Amino acid sequence for Hebeloma cylindrosporum glucosidase, Uniparc reference UPI00059A3BA8, Uniprot reference A0A0C2YJF2.

[0200] SEQ ID NO: 52: Amino acid sequence for Brenneria goodwinii glucosidase, Uniparc reference UPI0006579FEA, Uniprot reference A0A0G4JRR8.

[0201] SEQ ID NO: 53: Amino acid sequence for Aspergillus calidoustus glucosidase, Uniparc reference UPI00073C9E60, Uniprot reference A0A0U5GN20.

[0202] SEQ ID NO: 54: Amino acid sequence for Bacteroides finegoldii glucosidase, Uniparc reference UPI0006C6E0C3, Uniprot reference A0A174BEZ2.

[0203] SEQ ID NO: 55: Amino acid sequence for Altererythrobacter dongtanensis glucosidase, Uniparc reference UPI0008153D6D, Uniprot reference A0A1B2A943.

[0204] SEQ ID NO: 56: Amino acid sequence for uncultured Anaerotruncus sp. glucosidase, Uniparc reference UPI0008206E7B, Uniprot reference A0A1C6FWD9.

[0205] SEQ ID NO: 57: Amino acid sequence for Candidatus firestone glucosidase, Uniparc reference UPI0008AC0E17, Uniprot reference A0A1F5UJR6.

[0206] SEQ ID NO: 58: Amino acid sequence for Tenericutes bacterium glucosidase, Uniparc reference UPI0009C6C8E4, Uniprot reference A0A1V5HS22.

[0207] SEQ ID NO: 59: Amino acid sequence for Lentisphaerae bacterium glucosidase, Uniparc reference UPI0009CEE1AB, Uniprot reference A0A1V5VBL2.

[0208] SEQ ID NO: 60: Amino acid sequence for Planctomycetes bacterium glucosidase, Uniparc reference UPI0009CAAB0A, Uniprot reference A0A1V6FZ47.

[0209] SEQ ID NO: 61: Amino acid sequence for Pseudallescheria apiosperma glucosidase, Uniparc reference UPI0004DD62AC, Uniprot reference A0A084G332.

[0210] SEQ ID NO: 62: Amino acid sequence for Nonlabens sediminis glucosidase, Uniparc reference UPI000507F00A, Uniprot reference A0A090Q4N8.

[0211] SEQ ID NO: 63: Amino acid sequence for Gynuella sunshinyii glucosidase, Uniparc reference UPI0005CC42CA, Uniprot reference A0A0C5VDU3.

[0212] SEQ ID NO: 64: Amino acid sequence for Verticillium longisporum glucosidase, Uniparc reference UPI00063E4005, Uniprot reference A0A0G4N9Q7.

[0213] SEQ ID NO: 65: Amino acid sequence for Cellulomonas sp. glucosidase, Uniparc reference UPI00073C6CD3, Uniprot reference A0A0V8TAB5.

[0214] SEQ ID NO: 66: Amino acid sequence for Hungatella hathewayi glucosidase, Uniparc reference UPI0006C024BB, Uniprot reference A0A174FBX7.

[0215] SEQ ID NO: 67: Amino acid sequence for Mesorhizobium sp. glucosidase, Uniparc reference UPI000688E74D, Uniprot reference A0A1C2DG64.

[0216] SEQ ID NO: 68: Amino acid sequence for Clostridium sp. glucosidase, Uniparc reference UPI000822FAB7, Uniprot reference A0A1C6GRT5.

[0217] SEQ ID NO: 69: Amino acid sequence for Chlamydiales bacterium glucosidase, Uniparc reference UPI0009284E74, Uniprot reference A0A1M3CSY6.

[0218] SEQ ID NO: 70: Amino acid sequence for Spirochaetes bacterium glucosidase, Uniparc reference UPI0009CB461D, Uniprot reference A0A1V5HUX3.

[0219] SEQ ID NO: 71: Amino acid sequence for bacterium glucosidase, Uniparc reference UPI0009C92D6A, Uniprot reference A0A1V5VHB9.

[0220] SEQ ID NO: 72: Amino acid sequence for Thermotogae bacterium glucosidase, Uniparc reference UPI0009D12B86, Uniprot reference A0A1V6H2W5.

[0221] SEQ ID NO: 73: Amino acid sequence for Pseudallescheria apiosperma glucosidase, Uniparc reference UPI0004DCBED3, Uniprot reference A0A084GGE2.

[0222] SEQ ID NO: 74: Amino acid sequence for Algibacter lectus glucosidase, Uniparc reference UPI00050E0BE3, Uniprot reference A0A090VF17.

[0223] SEQ ID NO: 75: Amino acid sequence for Paxillus involutus glucosidase, Uniparc reference UPI0005B075C8, Uniprot reference A0A0C9TWP5.

[0224] SEQ ID NO: 76: Amino acid sequence for Verticillium longisporum glucosidase, Uniparc reference UPI00063DF006, Uniprot reference A0A0G4NA55.

[0225] SEQ ID NO: 77: Amino acid sequence for Mucilaginibacter gotjawali glucosidase, Uniparc reference UPI00076F8EA4, Uniprot reference A0A110B1H1.

[0226] SEQ ID NO: 78: Amino acid sequence for Bacteroides uniformis glucosidase, Uniparc reference UPI0006C0FFEF, Uniprot reference A0A174IWW4.

[0227] SEQ ID NO: 79: Amino acid sequence for Coprococcus sp. glucosidase, Uniparc reference UPI0008222B77, Uniprot reference A0A1C5W0N6.

[0228] SEQ ID NO: 80: Amino acid sequence for Blautia sp. glucosidase, Uniparc reference UPI0006C36823, Uniprot reference A0A1C6K2X5.

[0229] SEQ ID NO: 81: Amino acid sequence for Cellulomonas sp. glucosidase, Uniparc reference UPI00092B3CD4, Uniprot reference A0A1M3ELH4.

[0230] SEQ ID NO: 82: Amino acid sequence for bacterium glucosidase, Uniparc reference UPI0009C85428, Uniprot reference A0A1V5J984.

[0231] SEQ ID NO: 83: Amino acid sequence for Spirochaetes bacterium glucosidase, Uniparc reference UPI0009CA1D5C, Uniprot reference A0A1V5WHQ7.

[0232] SEQ ID NO: 84: Amino acid sequence for Tenericutes bacterium glucosidase, Uniparc reference UPI0009C7041F, Uniprot reference A0A1V6IJ57.

[0233] SEQ ID NO: 85: Amino acid sequence for Flavobacterium gilvum glucosidase, Uniparc reference UPI0004E2A41F, Uniprot reference A0A085EG29.

[0234] SEQ ID NO: 86: Amino acid sequence for Algibacter lectus glucosidase, Uniparc reference UPI0005102470, Uniprot reference A0A090WWZ1.

[0235] SEQ ID NO: 87: Amino acid sequence for Hydnomerulius pinastri glucosidase, Uniparc reference UPI0005B0F0AE, Uniprot reference A0A0C9WDY0.

[0236] SEQ ID NO: 88: Amino acid sequence for Nocardia farcinica glucosidase, Uniparc reference UPI00065C2666, Uniprot reference A0A0H5NWN2.

[0237] SEQ ID NO: 89: Amino acid sequence for Bacteroides cellulosilyticus glucosidase, Uniparc reference UPI000760375B, Uniprot reference A0A125MG18.

[0238] SEQ ID NO: 90: Amino acid sequence for Fonsecaea erecta glucosidase, Uniparc reference UPI0007DF4250, Uniprot reference A0A178ZBN2.

[0239] SEQ ID NO: 91: Amino acid sequence for Bacteroides sp. glucosidase, Uniparc reference UPI0008209852, Uniprot reference A0A1C5W9N9.

[0240] SEQ ID NO: 92: Amino acid sequence for Tannerella forsythia glucosidase, Uniparc reference UPI00086C34C1, Uniprot reference A0A1D3UGH8.

[0241] SEQ ID NO: 93: Amino acid sequence for Microbacterium esteraromaticum glucosidase, Uniparc reference UPI00097E83BB, Uniprot reference A0A1R4K192.

[0242] SEQ ID NO: 94: Amino acid sequence for bacterium glucosidase, Uniparc reference UPI0009D255E5, Uniprot reference A0A1V5LJK9.

[0243] SEQ ID NO: 95: Amino acid sequence for Candidatus hydrogenedentes glucosidase, Uniparc reference UPI0009C5A3CF, Uniprot reference A0A1V5Z2L2.

[0244] SEQ ID NO: 96: Amino acid sequence for Bacteroidetes bacterium glucosidase, Uniparc reference UPI0009C55799, Uniprot reference A0A1V6J4J8.

[0245] SEQ ID NO: 97: Amino acid sequence for Penicillium solitum glucosidase, Uniparc reference UPI0009D4067F, Uniprot reference A0A1V6RQ41.

[0246] SEQ ID NO: 98: Amino acid sequence for Weissella soli glucosidase, Uniparc reference UPI0008737AA2, Uniprot reference A0A288Q812.

[0247] SEQ ID NO: 99: Amino acid sequence for Acetatifactor muris glucosidase, Uniparc reference UPI000CAC57D4, Uniprot reference A0A2K4ZN91.

[0248] SEQ ID NO: 100: Amino acid sequence for Corynespora cassiicola glucosidase, Uniparc reference UPI000D237A4A, Uniprot reference A0A2T2NYD4.

[0249] SEQ ID NO: 101: Amino acid sequence for Meira miltonrushii glucosidase, Uniparc reference UPI000D77C91D, Uniprot reference A0A316V6M3.

[0250] SEQ ID NO: 102: Amino acid sequence for Bacteroides fragilis glucosidase, Uniparc reference UPI00004E1F76, Uniprot reference A0A380YVC7.

[0251] SEQ ID NO: 103: Amino acid sequence for Malassezia restricta glucosidase, Uniparc reference UPI000F0C30E8, Uniprot reference A0A3G2SB79.

[0252] SEQ ID NO: 104: Amino acid sequence for Fusarium euwallaceae glucosidase, Uniparc reference UPI000FFFEFB2, Uniprot reference A0A430LYA2.

[0253] SEQ ID NO: 105: Amino acid sequence for Psathyrella aberdarensis glucosidase, Uniparc reference UPI0010251887, Uniprot reference A0A4Q2E070.

[0254] SEQ ID NO: 106: Amino acid sequence for Aeromonas hydrophila glucosidase, Uniparc reference UPI0000E69509, Uniprot reference A0KLP6.

[0255] SEQ ID NO: 107: Amino acid sequence for Saccharopolyspora erythraea glucosidase, Uniparc reference UPI00000B86CB, Uniprot reference A4F7P9.

[0256] SEQ ID NO: 108: Amino acid sequence for Streptomyces sviceus glucosidase, Uniparc reference UPI000180240E, Uniprot reference B51181.

[0257] SEQ ID NO: 109: Amino acid sequence for Naematelia encephala glucosidase, Uniparc reference UPI000A250F78, Uniprot reference A0A1Y2AWB7.

[0258] SEQ ID NO: 110: Amino acid sequence for Hartmannibacter diazotrophicus glucosidase, Uniparc reference UPI000C0221F1, Uniprot reference A0A2C9D612.

[0259] SEQ ID NO: 111: Amino acid sequence for Pontimonas salivibrio glucosidase, Uniparc reference UPI000CEB5AB1, Uniprot reference A0A2L2BPE2.

[0260] SEQ ID NO: 112: Amino acid sequence for Cadophora sp. glucosidase, Uniparc reference UPI000D5B9C38, Uniprot reference A0A2V1CH24.

[0261] SEQ ID NO: 113: Amino acid sequence for Meira miltonrushii glucosidase, Uniparc reference UPI000D779558, Uniprot reference A0A316V8S0.

[0262] SEQ ID NO: 114: Amino acid sequence for Monilinia fructigena glucosidase, Uniparc reference UPI000DC42E3D, Uniprot reference A0A395IJW4.

[0263] SEQ ID NO: 115: Amino acid sequence for Hortaea werneckii glucosidase, Uniparc reference UPI000F3E476C, Uniprot reference A0A3M6XGS0.

[0264] SEQ ID NO: 116: Amino acid sequence for Streptomyces netropsis glucosidase, Uniparc reference UPI00101460D7, Uniprot reference A0A445N7U0.

[0265] SEQ ID NO: 117: Amino acid sequence for Aureobasidium pullulans glucosidase, Uniparc reference UPI001139C6A8, Uniprot reference A0A4S9IFI0.

[0266] SEQ ID NO: 118: Amino acid sequence for Aspergillus clavatus glucosidase, Uniparc reference UPI0000EA5CFF, Uniprot reference A1CTN9.

[0267] SEQ ID NO: 119: Amino acid sequence for Clavibacter michiganensis glucosidase, Uniparc reference UPI0001523037, Uniprot reference A5CT94.

[0268] SEQ ID NO: 120: Amino acid sequence for Penicillium rubens glucosidase, Uniparc reference UPI0001831CF5, Uniprot reference B6H7R5.

[0269] SEQ ID NO: 121: Amino acid sequence for Lachnoclostridium sp. glucosidase, Uniparc reference UPI000B365547, Uniprot reference A0A1Y4NTL9.

[0270] SEQ ID NO: 122: Amino acid sequence for Rhodobacteraceae bacterium glucosidase, Uniparc reference UPI000C09BF88, Uniprot reference A0A2D5IXB9.

[0271] SEQ ID NO: 123: Amino acid sequence for Bacteroides fragilis glucosidase, Uniparc reference UPI0004B5EEF2, Uniprot reference A0A2M9UUC4.

[0272] SEQ ID NO: 124: Amino acid sequence for Aspergillus indologenus glucosidase, Uniparc reference UPI000D7FE1ED, Uniprot reference A0A2V51Y78.

[0273] SEQ ID NO: 125: Amino acid sequence for Acaromyces ingoldii glucosidase, Uniparc reference UPI000D802B25, Uniprot reference A0A316YR39.

[0274] SEQ ID NO: 126: Amino acid sequence for Monilinia fructigena glucosidase, Uniparc reference UPI000DC60823, Uniprot reference A0A395J1U5.

[0275] SEQ ID NO: 127: Amino acid sequence for Paenibacillus xylanexedens glucosidase, Uniparc reference UPI000F52D773, Uniprot reference A0A3N6CA02.

[0276] SEQ ID NO: 128: Amino acid sequence for Actinomyces howellii glucosidase, Uniparc reference UPI000F6DAAAE, Uniprot reference A0A448HIG0.

[0277] SEQ ID NO: 129: Amino acid sequence for Friedmanniomyces endolithicus glucosidase, Uniparc reference UPI00113D19DF, Uniprot reference A0A4V5N914.

[0278] SEQ ID NO: 130: Amino acid sequence for Neosartorya fischeri glucosidase, Uniparc reference UPI0000EA8672, Uniprot reference A1DNS0.

[0279] SEQ ID NO: 131: Amino acid sequence for Pseudomonas aeruginosa glucosidase, Uniparc reference UPI0000D7314B, Uniprot reference A6V4K6.

[0280] SEQ ID NO: 132: Amino acid sequence for Talaromyces stipitatus glucosidase, Uniparc reference UPI00018E7266, Uniprot reference B8MF24.

[0281] SEQ ID NO: 133: Amino acid sequence for Aquimixticola soesokkakensis glucosidase, Uniparc reference UPI000A1A5FD7, Uniprot reference A0A1Y5RVF8.

[0282] SEQ ID NO: 134: Amino acid sequence for Rhodobacterales bacterium glucosidase, Uniparc reference UPI000C98D37C, Uniprot reference A0A2D9YGV1.

[0283] SEQ ID NO: 135: Amino acid sequence for Methylorubrum extorquens glucosidase, Uniparc reference UPI0006F9793E, Uniprot reference A0A2N9AS40.

[0284] SEQ ID NO: 136: Amino acid sequence for Clostridium perfringens glucosidase, Uniparc reference UPI00000CF7C8, Uniprot reference A0A2X2YBP0.

[0285] SEQ ID NO: 137: Amino acid sequence for Acholeplasmatales bacterium glucosidase, Uniparc reference UPI0008AEBEA3, Uniprot reference A0A348NID6.

[0286] SEQ ID NO: 138: Amino acid sequence for Bacteroidetes bacterium glucosidase, Uniparc reference UPI000EC3C979, Uniprot reference A0A3B8VHE8.

[0287] SEQ ID NO: 139: Amino acid sequence for Clostridium carnis glucosidase, Uniparc reference UPI000F637E38, Uniprot reference A0A3P6K8E8.

[0288] SEQ ID NO: 140: Amino acid sequence for Mycolicibacterium flavescens glucosidase, Uniparc reference UPI000B93B5C9, Uniprot reference A0A448HNB5.

[0289] SEQ ID NO: 141: Amino acid sequence for Streptococcus gallolyticus glucosidase, Uniparc reference UPI000F6EFA71, Uniprot reference A0A4V6LJ94.

[0290] SEQ ID NO: 142: Amino acid sequence for Yersinia enterocolitica glucosidase, Uniparc reference UPI0000EB54CC, Uniprot reference A1JNB7.

[0291] SEQ ID NO: 143: Amino acid sequence for Anaeromyxobacter sp. glucosidase, Uniparc reference UPI0000ED8A80, Uniprot reference A7HFG4.

[0292] SEQ ID NO: 144: Amino acid sequence for Talaromyces stipitatus glucosidase, Uniparc reference UPI00018E7D70, Uniprot reference B8MK55.

[0293] SEQ ID NO: 145: Amino acid sequence for Hortaea werneckii glucosidase, Uniparc reference UPI000A2E3FAA, Uniprot reference A0A1Z5SL14.

[0294] SEQ ID NO: 146: Amino acid sequence for Micavibrio sp. glucosidase, Uniparc reference UPI000C529025, Uniprot reference A0A2E2Q8X2.

[0295] SEQ ID NO: 147: Amino acid sequence for Acidobacteriia bacterium glucosidase, Uniparc reference UPI0000CE6B996, Uniprot reference A0A2N9MBS0.

[0296] SEQ ID NO: 148: Amino acid sequence for Corynebacterium jeikeium glucosidase, Uniparc reference UPI000DA3A972, Uniprot reference A0A2X4T570.

[0297] SEQ ID NO: 149: Amino acid sequence for Clostridiaceae bacterium glucosidase, Uniparc reference UPI000E8D37A1, Uniprot reference A0A353PZH8.

[0298] SEQ ID NO: 150: Amino acid sequence for Anaerolineaceae bacterium glucosidase, Uniparc reference UPI000748C096, Uniprot reference A0A3B9PA35.

[0299] SEQ ID NO: 151: Amino acid sequence for Gymnopilus dilepis glucosidase, Uniparc reference UPI000FF41956, Uniprot reference A0A409WSY0.

[0300] SEQ ID NO: 152: Amino acid sequence for Kocuria rosea glucosidase, Uniparc reference UPI000F7105D4, Uniprot reference A0A448R8N0.

[0301] SEQ ID NO: 153: Amino acid sequence for Teredinibacter sp. glucosidase, Uniparc reference UPI0011696FAB, Uniprot reference A0A509DWZ3.

[0302] SEQ ID NO: 154: Amino acid sequence for Aspergillus niger glucosidase, Uniparc reference UPI0000EFB564, Uniprot reference A2QS42.

[0303] SEQ ID NO: 155: Amino acid sequence for Laccaria bicolor glucosidase, Uniparc reference UPI000164423D, Uniprot reference B0D734.

[0304] SEQ ID NO: 156: Amino acid sequence for Pedosphaera parvula glucosidase, Uniparc reference UPI00017357F6, Uniprot reference B9XH33.

[0305] SEQ ID NO: 157: Amino acid sequence for Megamonas hypermegale glucosidase, Uniparc reference UPI00042469F6, Uniprot reference A0A239TGH2.

[0306] SEQ ID NO: 158: Amino acid sequence for Armillaria gallica glucosidase, Uniparc reference UPI000BC209C3, Uniprot reference A0A2H3E300.

[0307] SEQ ID NO: 159: Amino acid sequence for Micromonospora sp. glucosidase, Uniparc reference UPI000D2EAE87, Uniprot reference A0A2P8AV03.

[0308] SEQ ID NO: 160: Amino acid sequence for Klebsiella oxytoca glucosidase, Uniparc reference UPI000DA286EE, Uniprot reference A0A2X5CJC5.

[0309] SEQ ID NO: 161: Amino acid sequence for Candidatus ozemobacter glucosidase, Uniparc reference UPI000DFAEF6C, Uniprot reference A0A367ZIH8.

[0310] SEQ ID NO: 162: Amino acid sequence for Coleophoma crateriformis glucosidase, Uniparc reference UPI000E399EE8, Uniprot reference A0A3D8R2C2.

[0311] SEQ ID NO: 163: Amino acid sequence for Apiotrichum porosum glucosidase, Uniparc reference UPI000FBC01E3, Uniprot reference A0A427XHS2.

[0312] SEQ ID NO: 164: Amino acid sequence for Acholeplasma hippikon glucosidase, Uniparc reference UPI00068E4E50, Uniprot reference A0A449BJ27.

[0313] SEQ ID NO: 165: Amino acid sequence for Streptomyces spectabilis glucosidase, Uniparc reference UPI001185F074, Uniprot reference A0A516RGT1.

[0314] SEQ ID NO: 166: Amino acid sequence for Aspergillus niger glucosidase, Uniparc reference UPI0000EFCED2, Uniprot reference A2R8G2.

[0315] SEQ ID NO: 167: Amino acid sequence for Xanthomonas campestris glucosidase, Uniparc reference UPI00000D8BFA, Uniprot reference B0RYA0.

[0316] SEQ ID NO: 168: Amino acid sequence for Lactobacillus paracasei glucosidase, Uniparc reference UPI00019C9CD7, Uniprot reference C2FDL2.

[0317] SEQ ID NO: 169: Amino acid sequence for Bifiguratus adelaidae glucosidase, Uniparc reference UPI000BC490A3, Uniprot reference A0A261XUH4.

[0318] SEQ ID NO: 170: Amino acid sequence for bacterium glucosidase, Uniparc reference UPI000CC31AE2, Uniprot reference A0A2H5YYA1.

[0319] SEQ ID NO: 171: Amino acid sequence for Actinomadura parvosata glucosidase, Uniparc reference UPI000D26C9C6, Uniprot reference A0A2P91Y35.

[0320] SEQ ID NO: 172: Amino acid sequence for Melissococcus plutonius glucosidase, Uniparc reference UPI00024F22B8, Uniprot reference A0A2Z5Y4P3.

[0321] SEQ ID NO: 173: Amino acid sequence for Enterococcus durans glucosidase, Uniparc reference UPI000E020871, Uniprot reference A0A377KJS3.

[0322] SEQ ID NO: 174: Amino acid sequence for Malassezia restricta glucosidase, Uniparc reference UPI000DD17A7D, Uniprot reference A0A3G2S2J6.

[0323] SEQ ID NO: 175: Amino acid sequence for Apiotrichum porosum glucosidase, Uniparc reference UPI000FA2DE87, Uniprot reference A0A427XZQ0.

[0324] SEQ ID NO: 176: Amino acid sequence for Streptomonospora sp. glucosidase, Uniparc reference UPI0010355193, Uniprot reference A0A4P6PWP5.

[0325] SEQ ID NO: 177: Amino acid sequence for Lactobacillus gasseri glucosidase, Uniparc reference UPI00119640A1, Uniprot reference A0A558LH47.

[0326] SEQ ID NO: 178: Amino acid sequence for Aspergillus niger glucosidase, Uniparc reference UPI0000EFD0AA, Uniprot reference A2RAJ1.

[0327] SEQ ID NO: 179: Amino acid sequence for Neosartorya fumigata glucosidase, Uniparc reference UPI000170BF91, Uniprot reference B0XXG1.

[0328] SEQ ID NO: 180: Amino acid sequence for Eubacterium eligens glucosidase, Uniparc reference UPI0001A5B234, Uniprot reference C4Z6T5.

[0329] SEQ ID NO: 181: Amino acid sequence for Bifiguratus adelaidae glucosidase, Uniparc reference UPI000BC64A75, Uniprot reference A0A261XVM8.

[0330] SEQ ID NO: 182: Amino acid sequence for bacterium glucosidase, Uniparc reference UPI000CAA854E, Uniprot reference A0A2H6EX57.

[0331] SEQ ID NO: 183: Amino acid sequence for Corynespora cassiicola glucosidase, Uniparc reference UPI000D24EC2C, Uniprot reference A0A2T2N5H9.

[0332] SEQ ID NO: 184: Amino acid sequence for Pseudomicrostroma glucosiphilum glucosidase, Uniparc reference UPI000D77A0B7, Uniprot reference A0A316UK83.

[0333] SEQ ID NO: 185: Amino acid sequence for Staphylococcus saprophyticus glucosidase, Uniparc reference UPI0002DAAFCC, Uniprot reference A0A380HD31.

[0334] SEQ ID NO: 186: Amino acid sequence for Malassezia restricta glucosidase, Uniparc reference UPI000F0C6B8E, Uniprot reference A0A3G2S932.

[0335] SEQ ID NO: 187: Amino acid sequence for Saitozyma podzolica glucosidase, Uniparc reference UPI000FBA70E2, Uniprot reference A0A427YCL1.

[0336] SEQ ID NO: 188: Amino acid sequence for Tremella mesenterica glucosidase, Uniparc reference UPI00102822B4, Uniprot reference A0A4Q1BRC6.

[0337] SEQ ID NO: 189: Amino acid sequence for Arthrobacter sp. glucosidase, Uniparc reference UPI0000527506, Uniprot reference A0JZ86.

[0338] SEQ ID NO: 190: Amino acid sequence for Scheffersomyces stipitis glucosidase, Uniparc reference UPI000157388C, Uniprot reference A3LRB0.

[0339] SEQ ID NO: 191: Amino acid sequence for Leptothrix cholodnii glucosidase, Uniparc reference UPI0001712E02, Uniprot reference B1XZK8.

[0340] SEQ ID NO: 192: Amino acid sequence for Thauera sp. glucosidase, Uniparc reference UPI000166883C, Uniprot reference C4ZLL7.

[0341] SEQ ID NO: 193: Amino acid sequence for Kosmotoga olearia glucosidase, Uniparc reference UPI00018494AB, Uniprot reference C5CDW5.

[0342] SEQ ID NO: 194: Amino acid sequence for Roseburia intestinalis glucosidase, Uniparc reference UPI0001CD671F, Uniprot reference D4L3Y2.

[0343] SEQ ID NO: 195: Amino acid sequence for Streptococcus equinus glucosidase, Uniparc reference UPI0001E0DC00, Uniprot reference E0PDF8.

[0344] SEQ ID NO: 196: Amino acid sequence for Streptococcus cristatus glucosidase, Uniparc reference UPI0001F80C4B, Uniprot reference E8JUK5.

[0345] SEQ ID NO: 197: Amino acid sequence for Cellulosilyticum lentocellum glucosidase, Uniparc reference UPI0001D2DBBC, Uniprot reference F2JLH3.

[0346] SEQ ID NO: 198: Amino acid sequence for Streptococcus gallolyticus glucosidase, Uniparc reference UPI0001C48657, Uniprot reference F5WYI5.

[0347] SEQ ID NO: 199: Amino acid sequence for Ketogulonicigenium vulgare glucosidase, Uniparc reference UPI00021D443A, Uniprot reference F9Y8Z7.

[0348] SEQ ID NO: 200: Amino acid sequence for Spathaspora passalidarum glucosidase, Uniparc reference UPI000228255D, Uniprot reference G3AGX1.

[0349] SEQ ID NO: 201: Amino acid sequence for Niastella koreensis glucosidase, Uniparc reference UPI00023F6F5F, Uniprot reference G8T9J3.

[0350] SEQ ID NO: 202: Amino acid sequence for Cellvibrio sp. glucosidase, Uniparc reference UPI000260108C, Uniprot reference I3IDC0.

[0351] SEQ ID NO: 203: Amino acid sequence for Flavobacterium sp. glucosidase, Uniparc reference UPI000272D1E0, Uniprot reference J1ACA0.

[0352] SEQ ID NO: 204: Amino acid sequence for Macrophomina phaseolina glucosidase, Uniparc reference UPI00028E7FE1, Uniprot reference K2S5D3.

[0353] SEQ ID NO: 205: Amino acid sequence for Kosmotoga olearia glucosidase, Uniparc reference UPI00018483A2, Uniprot reference C5CHI5.

[0354] SEQ ID NO: 206: Amino acid sequence for Blautia obeum glucosidase, Uniparc reference UPI0001CD5918, Uniprot reference D4LRF6.

[0355] SEQ ID NO: 207: Amino acid sequence for Bifidobacterium dentium glucosidase, Uniparc reference UPI0001E18CDA, Uniprot reference E0Q541.

[0356] SEQ ID NO: 208: Amino acid sequence for Anaerolinea thermophila glucosidase, Uniparc reference UPI0001F55F8F, Uniprot reference E8N5R8.

[0357] SEQ ID NO: 209: Amino acid sequence for Coriobacterium glomerans glucosidase, Uniparc reference UPI0002050DA2, Uniprot reference F2N7E4.

[0358] SEQ ID NO: 210: Amino acid sequence for Microlunatus phosphovorus glucosidase, Uniparc reference UPI000210C886, Uniprot reference F5XJQ3.

[0359] SEQ ID NO: 211: Amino acid sequence for Streptomyces sp. glucosidase, Uniparc reference UPI0001C18877, Uniprot reference G0Q1U8.

[0360] SEQ ID NO: 212: Amino acid sequence for Spathaspora passalidarum glucosidase, Uniparc reference UPI0002282B61, Uniprot reference G3AIV6.

[0361] SEQ ID NO: 213: Amino acid sequence for Glarea lozoyensis glucosidase, Uniparc reference UPI0002402F4A, Uniprot reference H0ET34.

[0362] SEQ ID NO: 214: Amino acid sequence for Glaciozyma antarctica glucosidase, Uniparc reference UPI0002633B36, Uniprot reference I3UJK0.

[0363] SEQ ID NO: 215: Amino acid sequence for Arthrobacter sp. glucosidase, Uniparc reference UPI00027DFD35, Uniprot reference J7LN00.

[0364] SEQ ID NO: 216: Amino acid sequence for Fusarium pseudograminearum glucosidase, Uniparc reference UPI00028D698C, Uniprot reference K3VMA9.

[0365] SEQ ID NO: 217: Amino acid sequence for Hypocrea rufa glucosidase, Uniparc reference UPI000006AA61, Uniprot reference C6GGC9.

[0366] SEQ ID NO: 218: Amino acid sequence for Ruminococcus torques glucosidase, Uniparc reference UPI0001CDA964, Uniprot reference D4M6W6.

[0367] SEQ ID NO: 219: Amino acid sequence for Bifidobacterium dentium glucosidase, Uniparc reference UPI0001E17370, Uniprot reference E0Q9Z7.

[0368] SEQ ID NO: 220: Amino acid sequence for Bacteroides salanitronis glucosidase, Uniparc reference UPI0001FC71F6, Uniprot reference F0R2D7.

[0369] SEQ ID NO: 221: Amino acid sequence for Bacteroides coprosuis glucosidase, Uniparc reference UPI00020E6D19, Uniprot reference F3ZQ40.

[0370] SEQ ID NO: 222: Amino acid sequence for Marinomonas posidonica glucosidase, Uniparc reference UPI00020D4CC6, Uniprot reference F6CWF6.

[0371] SEQ ID NO: 223: Amino acid sequence for Chaetomium thermophilum glucosidase, Uniparc reference UPI000227E8ED, Uniprot reference G0 SE64.

[0372] SEQ ID NO: 224: Amino acid sequence for Tetragenococcus halophilus glucosidase, Uniparc reference UPI00022B9A15, Uniprot reference G4L5K1.

[0373] SEQ ID NO: 225: Amino acid sequence for Paenibacillus sp. glucosidase, Uniparc reference UPI00024F0867, Uniprot reference H6CCZ8.

[0374] SEQ ID NO: 226: Amino acid sequence for Turneriella parva glucosidase, Uniparc reference UPI000265AA4B, Uniprot reference 14B8U7.

[0375] SEQ ID NO: 227: Amino acid sequence for Arthrobacter sp. glucosidase, Uniparc reference UPI00027DFDB2, Uniprot reference J7LQK9.

[0376] SEQ ID NO: 228: Amino acid sequence for Agaricus bisporus glucosidase, Uniparc reference UPI00029074B1, Uniprot reference K5W7V1.

[0377] SEQ ID NO: 229: Amino acid sequence for Nectria haematococca glucosidase, Uniparc reference UPI0001B67634, Uniprot reference C7YIP3.

[0378] SEQ ID NO: 230: Amino acid sequence for Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0001A25287, Uniprot reference D4VSZ0.

[0379] SEQ ID NO: 231: Amino acid sequence for Sediminispirochaeta smaragdinae glucosidase, Uniparc reference UPI0001DD9790, Uniprot reference E1R331.

[0380] SEQ ID NO: 232: Amino acid sequence for Deinococcus proteolyticus glucosidase, Uniparc reference UPI0001FC42E9, Uniprot reference F0RPV2.

[0381] SEQ ID NO: 233: Amino acid sequence for Sphingobacterium sp. glucosidase, Uniparc reference UPI0002033AOA, Uniprot reference F4C226.

[0382] SEQ ID NO: 234: Amino acid sequence for Sphingobium chlorophenolicum glucosidase, Uniparc reference UPI0001E54133, Uniprot reference F6ET40.

[0383] SEQ ID NO: 235: Amino acid sequence for Caloramator australicus glucosidase, Uniparc reference UPI00021CACC4, Uniprot reference G0V3V5.

[0384] SEQ ID NO: 236: Amino acid sequence for Commensalibacter intestini glucosidase, Uniparc reference UPI000230E3BF, Uniprot reference G6F370.

[0385] SEQ ID NO: 237: Amino acid sequence for Paenibacillus sp. glucosidase, Uniparc reference UPI00024F07AC, Uniprot reference H6CIT2.

[0386] SEQ ID NO: 238: Amino acid sequence for Nitrolancea hollandica glucosidase, Uniparc reference UPI0002638AF3, Uniprot reference I4EIA9.

[0387] SEQ ID NO: 239: Amino acid sequence for Cryptococcus neoformans glucosidase, Uniparc reference UPI000392C3ED, Uniprot reference J9VVK7.

[0388] SEQ ID NO: 240: Amino acid sequence for Acidipropionibacterium acidipropionici glucosidase, Uniparc reference UPI0002988588, Uniprot reference K7S596.

[0389] SEQ ID NO: 241: Amino acid sequence for Prevotella sp. glucosidase, Uniparc reference UPI0001B93465, Uniprot reference C9PT75.

[0390] SEQ ID NO: 242: Amino acid sequence for Rhodobacter capsulatus glucosidase, Uniparc reference UPI0001D08095, Uniprot reference D5ALU0.

[0391] SEQ ID NO: 243: Amino acid sequence for Stigmatella aurantiaca glucosidase, Uniparc reference UPI0001E74370, Uniprot reference E3FJ05.

[0392] SEQ ID NO: 244: Amino acid sequence for Sphaerochaeta globosa glucosidase, Uniparc reference UPI0002010060, Uniprot reference F0RVK3.

[0393] SEQ ID NO: 245: Amino acid sequence for Sphaerochaeta coccoides glucosidase, Uniparc reference UPI000207D78C, Uniprot reference F4GH96.

[0394] SEQ ID NO: 246: Amino acid sequence for Novosphingobium sp. glucosidase, Uniparc reference UPI00020EFBCB, Uniprot reference F6ICQ5.

[0395] SEQ ID NO: 247: Amino acid sequence for Arthrobotrys oligospora glucosidase, Uniparc reference UPI000225331F, Uniprot reference G1XH86.

[0396] SEQ ID NO: 248: Amino acid sequence for Lactococcus lactis glucosidase, Uniparc reference UPI00000C6907, Uniprot reference G6FFS4.

[0397] SEQ ID NO: 249: Amino acid sequence for Phaeospirillum molischianum glucosidase, Uniparc reference UPI000255313A, Uniprot reference H8FXH7.

[0398] SEQ ID NO: 250: Amino acid sequence for Modestobacter marinus glucosidase, Uniparc reference UPI0002609C68, Uniprot reference I4EW72.

[0399] SEQ ID NO: 251: Amino acid sequence for Saccharothrix espanaensis glucosidase, Uniparc reference UPI00028AC337, Uniprot reference K0K125.

[0400] SEQ ID NO: 252: Amino acid sequence for Cronobacter sakazakii glucosidase, Uniparc reference UPI00029BA293, Uniprot reference K8DAK0.

[0401] SEQ ID NO: 253: Amino acid sequence for Verticillium alfalfae glucosidase, Uniparc reference UPI0001BBDF1E, Uniprot reference C9SVX1.

[0402] SEQ ID NO: 254: Amino acid sequence for Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0001CCFD36, Uniprot reference D6CY10.

[0403] SEQ ID NO: 255: Amino acid sequence for Leadbetterella byssophila glucosidase, Uniparc reference UPI0001EBD98A, Uniprot reference E4RUR0.

[0404] SEQ ID NO: 256: Amino acid sequence for Sphaerochaeta globosa glucosidase, Uniparc reference UPI00020102A5, Uniprot reference F0RYB6.

[0405] SEQ ID NO: 257: Amino acid sequence for Sphaerochaeta coccoides glucosidase, Uniparc reference UPI000207D6A0, Uniprot reference F4GLH6.

[0406] SEQ ID NO: 258: Amino acid sequence for Haloplasma contractile glucosidase, Uniparc reference UPI000212252C, Uniprot reference F7Q0Y2.

[0407] SEQ ID NO: 259: Amino acid sequence for Nitrospirillum amazonense glucosidase, Uniparc reference UPI0002265447, Uniprot reference G1Y4C7.

[0408] SEQ ID NO: 260: Amino acid sequence for Azospirillum brasilense glucosidase, Uniparc reference UPI00023420B8, Uniprot reference G8AWD9.

[0409] SEQ ID NO: 261: Amino acid sequence for Phaeospirillum molischianum glucosidase, Uniparc reference UPI000255314F, Uniprot reference H8FXJ8.

[0410] SEQ ID NO: 262: Amino acid sequence for Modestobacter marinus glucosidase, Uniparc reference UPI000260A2FA, Uniprot reference I4EYD5.

[0411] SEQ ID NO: 263: Amino acid sequence for Wickerhamomyces ciferrii glucosidase, Uniparc reference UPI000283EB8D, Uniprot reference K0KVJ2.

[0412] SEQ ID NO: 264: Amino acid sequence for Gloeocapsa sp. glucosidase, Uniparc reference UPI0002A5D085, Uniprot reference K9XKL8.

[0413] SEQ ID NO: 265: Amino acid sequence for Sphaerobacter thermophilus glucosidase, Uniparc reference UPI0001A3BCB6, Uniprot reference D1C7U8.

[0414] SEQ ID NO: 266: Amino acid sequence for Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0001CCEF1F, Uniprot reference D6D4V2.

[0415] SEQ ID NO: 267: Amino acid sequence for Prevotella buccae glucosidase, Uniparc reference UPI0001F149E8, Uniprot reference E6K4W5.

[0416] SEQ ID NO: 268: Amino acid sequence for Grosmannia clavigera glucosidase, Uniparc reference UPI0001FF1101, Uniprot reference F0XBR0.

[0417] SEQ ID NO: 269: Amino acid sequence for Melampsora larici-populina glucosidase, Uniparc reference UPI00020F9774, Uniprot reference F4R4W2.

[0418] SEQ ID NO: 270: Amino acid sequence for Prevotella multisaccharivorax glucosidase, Uniparc reference UPI0002138E9C, Uniprot reference F8N7G1.

[0419] SEQ ID NO: 271: Amino acid sequence for Streptomyces zinciresistens glucosidase, Uniparc reference UPI0002255A63, Uniprot reference G2G8K4.

[0420] SEQ ID NO: 272: Amino acid sequence for Granulicella mallensis glucosidase, Uniparc reference UPI0001D9FC40, Uniprot reference G8NY42.

[0421] SEQ ID NO: 273: Amino acid sequence for Gibberella zeae glucosidase, Uniparc reference UPI00021F1FEE, Uniprot reference I1RH94.

[0422] SEQ ID NO: 274: Amino acid sequence for Modestobacter marinus glucosidase, Uniparc reference UPI00026090A2, Uniprot reference I4EYK6.

[0423] SEQ ID NO: 275: Amino acid sequence for Lactobacillus equicursoris glucosidase, Uniparc reference UPI0002869F55, Uniprot reference K0NRS8.

[0424] SEQ ID NO: 276: Amino acid sequence for Colletotrichum fructicola glucosidase, Uniparc reference UPI0002A93280, Uniprot reference L2F9W0.

[0425] SEQ ID NO: 277: Amino acid sequence for Streptosporangium roseum glucosidase, Uniparc reference UPI0001BF8AF6, Uniprot reference D2B261.

[0426] SEQ ID NO: 278: Amino acid sequence for Listeria grayi glucosidase, Uniparc reference UPI00019F252A, Uniprot reference D7UX19.

[0427] SEQ ID NO: 279: Amino acid sequence for Enterococcus italicus glucosidase, Uniparc reference UPI0001F11EFF, Uniprot reference E6LF07.

[0428] SEQ ID NO: 280: Amino acid sequence for Fluviicola taffensis glucosidase, Uniparc reference UPI000203D9EA, Uniprot reference F211T6.

[0429] SEQ ID NO: 281: Amino acid sequence for Shigella flexneri glucosidase, Uniparc reference UPI00020CA641, Uniprot reference F5N4W9.

[0430] SEQ ID NO: 282: Amino acid sequence for Actinomyces sp. glucosidase, Uniparc reference UPI0002189080, Uniprot reference F9EFW5.

[0431] SEQ ID NO: 283: Amino acid sequence for Verticillium dahliae glucosidase, Uniparc reference UPI00022EBE5A, Uniprot reference G2X5V6.

[0432] SEQ ID NO: 284: Amino acid sequence for Actinoplanes sp. glucosidase, Uniparc reference UPI00023ED5A8, Uniprot reference G8S0M0.

[0433] SEQ ID NO: 285: Amino acid sequence for Gibberella zeae glucosidase, Uniparc reference UPI000023EA29, Uniprot reference 11S320.

[0434] SEQ ID NO: 286: Amino acid sequence for Auricularia subglabra glucosidase, Uniparc reference UPI00027CE685, Uniprot reference J0WV06.

[0435] SEQ ID NO: 287: Amino acid sequence for Nitratireductor indicus glucosidase, Uniparc reference UPI00028E86D6, Uniprot reference K2N268.

[0436] SEQ ID NO: 288: Amino acid sequence for Thermoclostridium stercorarium glucosidase, Uniparc reference UPI0002AD999E, Uniprot reference L7VH66.

[0437] SEQ ID NO: 289: Amino acid sequence for Geobacillus sp. glucosidase, Uniparc reference UPI0002AF2DE3, Uniprot reference L8A1S2.

[0438] SEQ ID NO: 290: Amino acid sequence for uncultured bacterium glucosidase, Uniparc reference UPI000327C3CC, Uniprot reference M9ZC55.

[0439] SEQ ID NO: 291: Amino acid sequence for Burkholderia ambifaria glucosidase, Uniparc reference UPI000059ACE1, Uniprot reference Q0BAK2.

[0440] SEQ ID NO: 292: Amino acid sequence for Aspergillus oryzae glucosidase, Uniparc reference UPI0000676B8D, Uniprot reference Q2UIR4.

[0441] SEQ ID NO: 293: Amino acid sequence for Yersinia pseudotuberculosis glucosidase, Uniparc reference UPI00004269B5, Uniprot reference Q66DJ0.

[0442] SEQ ID NO: 294: Amino acid sequence for Wallemia ichthyophaga glucosidase, Uniparc reference UPI000331CC98, Uniprot reference R9AF64.

[0443] SEQ ID NO: 295: Amino acid sequence for Glarea lozoyensis glucosidase, Uniparc reference UPI0003522404, Uniprot reference S3DIM6.

[0444] SEQ ID NO: 296: Amino acid sequence for Moniliophthora roreri glucosidase, Uniparc reference UPI0003BF753A, Uniprot reference V2YU31.

[0445] SEQ ID NO: 297: Amino acid sequence for Zhouia amylolytica glucosidase, Uniparc reference UPI0003DBCB4F, Uniprot reference W2ULL3.

[0446] SEQ ID NO: 298: Amino acid sequence for Fusarium oxysporum glucosidase, Uniparc reference UPI0003F362FF, Uniprot reference W9NQQ3.

[0447] SEQ ID NO: 299: Amino acid sequence for Flavobacterium johnsoniae glucosidase, Uniparc reference UPI00006EA168, Uniprot reference A5FAA5.

[0448] SEQ ID NO: 300: Amino acid sequence for Oryza sativa glucosidase, Uniparc reference UPI00002394F5, Uniprot reference Q75193.

[0449] SEQ ID NO: 301: Amino acid sequence for Clostridium saccharoperbutylacetonicum glucosidase, Uniparc reference UPI0002B65681, Uniprot reference M1MCD1.

[0450] SEQ ID NO: 302: Amino acid sequence for Rhodococcus sp. glucosidase, Uniparc reference UPI0002D21DB1, Uniprot reference N1MBN6.

[0451] SEQ ID NO: 303: Amino acid sequence for Burkholderia ambifaria glucosidase, Uniparc reference UPI000059C0CD, Uniprot reference Q0BCV8.

[0452] SEQ ID NO: 304: Amino acid sequence for Xanthomonas campestris glucosidase, Uniparc reference UPI00005CE9E7, Uniprot reference Q3BVH7.

[0453] SEQ ID NO: 305: Amino acid sequence for Caulobacter vibrioides glucosidase, Uniparc reference UPI00000C7604, Uniprot reference Q9A6F8.

[0454] SEQ ID NO: 306: Amino acid sequence for Arcticibacter svalbardensis glucosidase, Uniparc reference UPI000338772A, Uniprot reference R9GRA7.

[0455] SEQ ID NO: 307: Amino acid sequence for Winogradskyella psychrotolerans glucosidase, Uniparc reference UPI00035A4604, Uniprot reference S7VQ28.

[0456] SEQ ID NO: 308: Amino acid sequence for Methyloglobulus morosus glucosidase, Uniparc reference UPI0003C4ED6A, Uniprot reference V5DXT8.

[0457] SEQ ID NO: 309: Amino acid sequence for Pestalotiopsis fici glucosidase, Uniparc reference UPI0003E05C04, Uniprot reference W3WV37.

[0458] SEQ ID NO: 310: Amino acid sequence for Capronia coronata glucosidase, Uniparc reference UPI000434E698, Uniprot reference W9YNR7.

[0459] SEQ ID NO: 311: Amino acid sequence for Aspergillus aculeatus glucosidase, Uniparc reference UPI00001268FD, Uniprot reference P48825.

[0460] SEQ ID NO: 312: Amino acid sequence for Thermotoga neapolitana glucosidase, Uniparc reference UPI0000DD5996, Uniprot reference Q0GC07.

[0461] SEQ ID NO: 313: Amino acid sequence for Clostridium saccharoperbutylacetonicum glucosidase, Uniparc reference UPI0002B666E8, Uniprot reference M1MJF4.

[0462] SEQ ID NO: 314: Amino acid sequence for Schizosaccharomyces pombe glucosidase, Uniparc reference UPI000006B001, Uniprot reference O74799.

[0463] SEQ ID NO: 315: Amino acid sequence for Phaeosphaeria nodorum glucosidase, Uniparc reference UPI000161BD2B, Uniprot reference Q0TXF6.

[0464] SEQ ID NO: 316: Amino acid sequence for Xylella fastidiosa Dixon glucosidase, Uniparc reference UPI00003806C8, Uniprot reference Q3RGJ3.

[0465] SEQ ID NO: 317: Amino acid sequence for Schizosaccharomyces pombe glucosidase, Uniparc reference UPI000006A330, Uniprot reference Q9P6J6.

[0466] SEQ ID NO: 318: Amino acid sequence for Arcticibacter svalbardensis glucosidase, Uniparc reference UPI000337E887, Uniprot reference R9GWD6.

[0467] SEQ ID NO: 319: Amino acid sequence for Colletotrichum gloeosporioides glucosidase, Uniparc reference UPI0003885717, Uniprot reference T0KJI7.

[0468] SEQ ID NO: 320: Amino acid sequence for uncultured bacterium glucosidase, Uniparc reference UPI0003C9E340, Uniprot reference V5R1E8.

[0469] SEQ ID NO: 321: Amino acid sequence for Xanthomonas arboricola glucosidase, Uniparc reference UPI0003E06A61, Uniprot reference W4S715.

[0470] SEQ ID NO: 322: Amino acid sequence for Fusarium oxysporum glucosidase, Uniparc reference UPI00021EC697, Uniprot reference X0A8X8.

[0471] SEQ ID NO: 323: Amino acid sequence for Paenibacillus polymyxa glucosidase, Uniparc reference UPI00001108DO, Uniprot reference P22073.

[0472] SEQ ID NO: 324: Amino acid sequence for Kluyveromyces marxianus glucosidase, Uniparc reference UPI0001BE5ADA, Uniprot reference D1GCC6.

[0473] SEQ ID NO: 325: Amino acid sequence for Ilumatobacter coccineus glucosidase, Uniparc reference UPI0002C04A25, Uniprot reference M5A594.

[0474] SEQ ID NO: 326: Amino acid sequence for Agrobacterium sp. glucosidase, Uniparc reference UPI0000126912, Uniprot reference P12614.

[0475] SEQ ID NO: 327: Amino acid sequence for Cytophaga hutchinsonii glucosidase, Uniparc reference UPI000038ECF7, Uniprot reference Q11P53.

[0476] SEQ ID NO: 328: Amino acid sequence for Thermobifida fusca glucosidase, Uniparc reference UPI00003C5CA4, Uniprot reference Q47PF5.

[0477] SEQ ID NO: 329: Amino acid sequence for Botryotinia fuckeliana glucosidase, Uniparc reference UPI0000069E5E, Uniprot reference Q9UVJ6.

[0478] SEQ ID NO: 330: Amino acid sequence for Agarivorans albus glucosidase, Uniparc reference UPI00033994D2, Uniprot reference R9PTK2.

[0479] SEQ ID NO: 331: Amino acid sequence for Enterococcus sp. glucosidase, Uniparc reference UPI00038B7639, Uniprot reference T0UDU2.

[0480] SEQ ID NO: 332: Amino acid sequence for Salinispira pacifica glucosidase, Uniparc reference UPI0003D8A7DC, Uniprot reference V5WKT4.

[0481] SEQ ID NO: 333: Amino acid sequence for Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0003ECF15E, Uniprot reference W6P696.

[0482] SEQ ID NO: 334: Amino acid sequence for Fusarium oxysporum glucosidase, Uniparc reference UPI000430029A, Uniprot reference X0AB46.

[0483] SEQ ID NO: 335: Amino acid sequence for Thermotoga maritima glucosidase, Uniparc reference UPI0000126906, Uniprot reference Q08638B.

[0484] SEQ ID NO: 336: Amino acid sequence for Neotermes koshunensis glucosidase, Uniparc reference UPI0000083EA8, Uniprot reference Q8T0W7.

[0485] SEQ ID NO: 337: Amino acid sequence for Thanatephorus cucumeris glucosidase, Uniparc reference UPI0002BF34B3, Uniprot reference M5CHG9.

[0486] SEQ ID NO: 338: Amino acid sequence for Hungateiclostridium thermocellum glucosidase, Uniparc reference UPI000053581D, Uniprot reference P14002.

[0487] SEQ ID NO: 339: Amino acid sequence for Koribacter versatilis glucosidase, Uniparc reference UPI0000D76A70, Uniprot reference Q1IJ89.

[0488] SEQ ID NO: 340: Amino acid sequence for Neosartorya fumigata glucosidase, Uniparc reference UPI00005203D8, Uniprot reference Q4WLX5.

[0489] SEQ ID NO: 341: Amino acid sequence for Amycolatopsis vancoresmycina glucosidase, Uniparc reference UPI00032DB8C1, Uniprot reference R1FKF0.

[0490] SEQ ID NO: 342: Amino acid sequence for Gibberella fujikuroi glucosidase, Uniparc reference UPI0003519C76, Uniprot reference S0EKU1.

[0491] SEQ ID NO: 343: Amino acid sequence for Bifidobacterium longum glucosidase, Uniparc reference UPI000390F839, Uniprot reference T212H5.

[0492] SEQ ID NO: 344: Amino acid sequence for uncultured bacterium glucosidase, Uniparc reference UPI0003DFF0DA, Uniprot reference W0FLD1.

[0493] SEQ ID NO: 345: Amino acid sequence for Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0002D3D994, Uniprot reference W6P9J9.

[0494] SEQ ID NO: 346: Amino acid sequence for Fusarium oxysporum glucosidase, Uniparc reference UPI00042F8016, Uniprot reference X0HA35.

[0495] SEQ ID NO: 347: Amino acid sequence for Sorghum bicolor glucosidase, Uniparc reference UPI00000A7F40, Uniprot reference Q41290.

[0496] SEQ ID NO: 348: Amino acid sequence for Clostridium cellulovorans glucosidase, Uniparc reference UPI000050B701, Uniprot reference Q53EH2.

[0497] SEQ ID NO: 349: Amino acid sequence for Dacryopinax primogenitus glucosidase, Uniparc reference UPI0002C29EAA, Uniprot reference M5G1U5.

[0498] SEQ ID NO: 350: Amino acid sequence for Rhizobium radiobacter glucosidase, Uniparc reference UPI0000126913, Uniprot reference P27034.

[0499] SEQ ID NO: 351: Amino acid sequence for Phanerochaete chrysosporium glucosidase, Uniparc reference UPI00006E0CC7, Uniprot reference Q25BW5A.

[0500] SEQ ID NO: 352: Amino acid sequence for Enterobacter agglomerans glucosidase, Uniparc reference UPI0000126905, Uniprot reference Q59437.

[0501] SEQ ID NO: 353: Amino acid sequence for Candidatus microthrix glucosidase, Uniparc reference UPI00033056C4, Uniprot reference R4YWU4.

[0502] SEQ ID NO: 354: Amino acid sequence for Chthonomonas calidirosea glucosidase, Uniparc reference UPI0003427F29, Uniprot reference S0ET37.

[0503] SEQ ID NO: 355: Amino acid sequence for Acholeplasma brassicae glucosidase, Uniparc reference UPI0003B04208, Uniprot reference U4KMR7.

[0504] SEQ ID NO: 356: Amino acid sequence for Chania multitudinisentens glucosidase, Uniparc reference UPI0003E13BD3, Uniprot reference W0LJ28.

[0505] SEQ ID NO: 357: Amino acid sequence for Gibberella moniliformis glucosidase, Uniparc reference UPI0003ECCF80, Uniprot reference W7MW98.

[0506] SEQ ID NO: 358: Amino acid sequence for Fusarium oxysporum glucosidase, Uniparc reference UPI0004305856, Uniprot reference X0I2U8.

[0507] SEQ ID NO: 359: Amino acid sequence for Phanerochaete chrysosporium glucosidase, Uniparc reference UPI00006E0CC7, Uniprot reference Q25BW5B.

[0508] SEQ ID NO: 360: Amino acid sequence for Secale cereale glucosidase, Uniparc reference UPI00000A7EFC, Uniprot reference Q9FYS3.

[0509] SEQ ID NO: 361: Amino acid sequence for Anoxybacillus gonensis glucosidase, Uniparc reference UPI0002C0273E, Uniprot reference M5QUM2.

[0510] SEQ ID NO: 362: Amino acid sequence for Thermotoga maritima glucosidase, Uniparc reference UPI0000126906, Uniprot reference Q08638A.

[0511] SEQ ID NO: 363: Amino acid sequence for Rhodospirillum rubrum glucosidase, Uniparc reference UPI00003C2ACC, Uniprot reference Q2RP51.

[0512] SEQ ID NO: 364: Amino acid sequence for Thermotoga neapolitana glucosidase, Uniparc reference UPI00000B9013, Uniprot reference Q60038.

[0513] SEQ ID NO: 365: Amino acid sequence for Candidatus microthrix glucosidase, Uniparc reference UPI00032F466E, Uniprot reference R4Z6M6.

[0514] SEQ ID NO: 366: Amino acid sequence for Ruminiclostridium cellobioparum glucosidase, Uniparc reference UPI00032874EC, Uniprot reference S0FPI8.

[0515] SEQ ID NO: 367: Amino acid sequence for Acholeplasma brassicae glucosidase, Uniparc reference UPI0003B0490C, Uniprot reference U4KMV4.

[0516] SEQ ID NO: 368: Amino acid sequence for Klebsiella pneumoniae glucosidase, Uniparc reference UPI0003DB6EFD, Uniprot reference W1BBP5.

[0517] SEQ ID NO: 369: Amino acid sequence for Enterobacter sp. glucosidase, Uniparc reference UPI0003ED0A8E, Uniprot reference W7P6Y1.

[0518] SEQ ID NO: 370: Amino acid sequence for Rhizoctonia solani glucosidase, Uniparc reference UPI00045BB507, Uniprot reference X8JI88.

[0519] SEQ ID NO: 371: Amino acid sequence for Homo sapiens glucosidase, Uniparc reference UPI0000072C73, Uniprot reference Q9H227.

[0520] SEQ ID NO: 372: Amino acid sequence for uncultured bacterium glucosidase, Uniparc reference UPI0000DD5304, Uniprot reference Q0GMU3.

[0521] SEQ ID NO: 373: Amino acid sequence for Thermoanaerobacter thermohydrosulfuricus glucosidase, Uniparc reference UPI0002CA9E4F, Uniprot reference M8CQD9.

[0522] SEQ ID NO: 374: Amino acid sequence for Stigmatella aurantiaca glucosidase, Uniparc reference UPI0000E297D8, Uniprot reference Q091M8.

[0523] SEQ ID NO: 375: Amino acid sequence for Burkholderia thailandensis glucosidase, Uniparc reference UPI00006676B1, Uniprot reference Q2T7C9.

[0524] SEQ ID NO: 376: Amino acid sequence for Yersinia pseudotuberculosis glucosidase, Uniparc reference UPI0000426E8F, Uniprot reference Q665S2.

[0525] SEQ ID NO: 377: Amino acid sequence for Togninia minima glucosidase, Uniparc reference UPI00032BF71F, Uniprot reference R8BQM8.

[0526] SEQ ID NO: 378: Amino acid sequence for Ophiostoma piceae glucosidase, Uniparc reference UPI00035213C2, Uniprot reference S3BVJ8.

[0527] SEQ ID NO: 379: Amino acid sequence for Pyronema omphalodes glucosidase, Uniparc reference UPI0003B117B7, Uniprot reference U4LIL3.

[0528] SEQ ID NO: 380: Amino acid sequence for Ogataea parapolymorpha glucosidase, Uniparc reference UPI0001F76D1F, Uniprot reference W1QIW0.

[0529] SEQ ID NO: 381: Amino acid sequence for Hymenobacter swuensis glucosidase, Uniparc reference UPI0003F2059F, Uniprot reference W8F6U0.

[0530] SEQ ID NO: 382: Amino acid sequence for Flavobacterium johnsoniae glucosidase, Uniparc reference UPI00006E8E1B, Uniprot reference A5FEF5.

[0531] SEQ ID NO: 383: Amino acid sequence for Paenibacillus polymyxa glucosidase, Uniparc reference UPI000012690B, Uniprot reference P22505.

[0532] SEQ ID NO: 384: Amino acid sequence for Oryza sativa glucosidase, Uniparc reference UPI000009D014, Uniprot reference Q8L7J2.

[0533] SEQ ID NO: 385: Amino acid sequence for Oryza sativa glucosidase, Uniparc reference UPI0000E580F2, Uniprot reference B8AVF0.

[0534] SEQ ID NO: 386: Amino acid sequence for Nannochloris glucosidase, Uniparc reference UPI000EA7F16F, Uniprot reference A0A452CSM4.

[0535] SEQ ID NO: 387: Amino acid sequence for Halothermothrix orenii glucosidase, Uniparc reference UPI00006AE508, Uniprot reference B8CYA8.

[0536] SEQ ID NO: 388: Amino acid sequence for Neurospora crassa glucosidase, Uniparc reference UPI000018B2B4, Uniprot reference Q7RWP2.

[0537] SEQ ID NO: 389: Amino acid sequence for Micrococcus antarcticus glucosidase, Uniparc reference UPI000192BB5F, Uniprot reference B9V8P5.

[0538] SEQ ID NO: 390: Amino acid sequence for Exiguobacterium antarcticum glucosidase, Uniparc reference UPI000285E79E, Uniprot reference K0A8J9.

[0539] SEQ ID NO: 391: Amino acid sequence for Thermus thermophilus glucosidase, Uniparc reference UPI00000BEB61, Uniprot reference Q9RA61.

[0540] SEQ ID NO: 392: Amino acid sequence for Trichoderma harzianum glucosidase, Uniparc reference UPI00078BF747, Uniprot reference A0A2T4AR08.

[0541] SEQ ID NO: 393: Amino acid sequence for Hypocrea jecorina glucosidase, Uniparc reference UPI000006AA61, Uniprot reference Q12715.

[0542] SEQ ID NO: 394: Amino acid sequence for Streptomyces sp. glucosidase, Uniparc reference UPI00000B411B, Uniprot reference Q59976.

[0543] SEQ ID NO: 395: Amino acid sequence for Streptococcus pyogenes glucosidase, Uniparc reference UPI00000C7E56, Uniprot reference Q99YP9.

[0544] SEQ ID NO: 396: Amino acid sequence for Trifolium repens glucosidase, Uniparc reference UPI000012691B, Uniprot reference P26205.

[0545] SEQ ID NO: 397: Amino acid sequence for Talaromyces emersonii glucosidase, Uniparc reference UPI000006C8FF, Uniprot reference Q8TGI8.

[0546] SEQ ID NO: 398: Amino acid sequence for Hungateiclostridium thermocellum glucosidase, Uniparc reference UPI0000126903, Uniprot reference P26208.

[0547] SEQ ID NO: 399: Amino acid sequence for Lactobacillus plantarum glucosidase, Uniparc reference UPI000219FE3E, Uniprot reference F9ULH8.

[0548] SEQ ID NO: 400: Amino acid sequence for Agrobacterium tumefaciens glucosidase, Uniparc reference UPI0003F2033A, Uniprot reference A0A2I4PGZ0.

[0549] SEQ ID NO: 401: Codon optimised DNA encoding 6×His Cyberlindnera fabianii glucosidase, Uniparc reference UPI00049B1A8C, Uniprot reference A0A061B3J2.

[0550] SEQ ID NO: 402: Codon optimised DNA encoding 6×His Flavobacterium gilvum glucosidase, Uniparc reference UPI0004E3EF7B, Uniprot reference A0A085EII0.

[0551] SEQ ID NO: 403: Codon optimised DNA encoding 6×His Algibacter lectus glucosidase, Uniparc reference UPI00050EE490, Uniprot reference A0A090X649.

[0552] SEQ ID NO: 404: Codon optimised DNA encoding 6×His Microbacterium azadirachtae glucosidase, Uniparc reference UPI0005ECB51E, Uniprot reference A0A0F0LB94.

[0553] SEQ ID NO: 405: Codon optimised DNA encoding 6×His Actinobacteria bacterium glucosidase, Uniparc reference UPI0006588DAD, Uniprot reference A0A0J0UT37.

[0554] SEQ ID NO: 406: Codon optimised DNA encoding 6×His Chloroflexi bacterium glucosidase, Uniparc reference UPI0007968552, Uniprot reference A0A136KWB3.

[0555] SEQ ID NO: 407: Codon optimised DNA encoding 6×His Komagataeibacter rhaeticus glucosidase, Uniparc reference UPI0002080410, Uniprot reference A0A181C809.

[0556] SEQ ID NO: 408: Codon optimised DNA encoding 6×His Bacteroides sp. glucosidase, Uniparc reference UPI0008211 BFC, Uniprot reference A0A1C5WEL8.

[0557] SEQ ID NO: 409: Codon optimised DNA encoding 6×His Streptomyces rubrolavendulae glucosidase, Uniparc reference UPI00085A2BD0, Uniprot reference A0A1 D8FZW3.

[0558] SEQ ID NO: 410: Codon optimised DNA encoding 6×His Clostridium roseum glucosidase, Uniparc reference UPI00098C60F6, Uniprot reference A0A1S8KYM5.

[0559] SEQ ID NO: 411: Codon optimised DNA encoding 6×His uncultured bacterium glucosidase, Uniparc reference UPI0009CE0D4C, Uniprot reference A0A1V5M6V6.

[0560] SEQ ID NO: 412: Codon optimised DNA encoding 6×His Firmicutes bacterium glucosidase, Uniparc reference UPI0009D4127D, Uniprot reference A0A1V6AN95.

[0561] SEQ ID NO: 413: Codon optimised DNA encoding 6×His Anthracocystis flocculosa glucosidase, Uniparc reference UPI00045601AB, Uniprot reference A0A061H1Z3.

[0562] SEQ ID NO: 414: Codon optimised DNA encoding 6×His Bifidobacterium boum glucosidase, Uniparc reference UPI0004FF77C7, Uniprot reference A0A086ZKU2.

[0563] SEQ ID NO: 415: Codon optimised DNA encoding 6×His Jejuia pallidilutea glucosidase, Uniparc reference UPI00051 EDBDE, Uniprot reference A0A098LTR2.

[0564] SEQ ID NO: 416: Codon optimised DNA encoding 6×His Ceratocystis fimbriata glucosidase, Uniparc reference UPI00062105AB, Uniprot reference A0A0F8B2B0.

[0565] SEQ ID NO: 417: Codon optimised DNA encoding 6×His Actinobacteria bacterium glucosidase, Uniparc reference UPI0006583AB1, Uniprot reference A0A0J0UVW7.

[0566] SEQ ID NO: 418: Codon optimised DNA encoding 6×His Rhodococcus sp. glucosidase, Uniparc reference UPI0007AABFAD, Uniprot reference A0A143QAX3.

[0567] SEQ ID NO: 419: Codon optimised DNA encoding 6×His Valsa mali glucosidase, Uniparc reference UPI0007F2D02D, Uniprot reference A0A194VF47.

[0568] SEQ ID NO: 420: Codon optimised DNA encoding 6×His uncultured Bacteroides sp. glucosidase, Uniparc reference UPI000821004D, Uniprot reference A0A1C5WS14.

[0569] SEQ ID NO: 421: Codon optimised DNA encoding 6×His Eisenbergiella tayi glucosidase, Uniparc reference UPI00084089B2, Uniprot reference A0A1E3ALT2.

[0570] SEQ ID NO: 422: Codon optimised DNA encoding 6×His Streptomyces sp. glucosidase, Uniparc reference UPI000978E914, Uniprot reference A0A1V2MYI4.

[0571] SEQ ID NO: 423: Codon optimised DNA encoding 6×His Firmicutes bacterium glucosidase, Uniparc reference UPI0009CBF21C, Uniprot reference A0A1V5MH90.

[0572] SEQ ID NO: 424: Codon optimised DNA encoding 6×His Tenericutes bacterium glucosidase, Uniparc reference UPI0009D5B1F0, Uniprot reference A0A1V6BAK3.

[0573] SEQ ID NO: 425: Codon optimised DNA encoding 6×His Gluconobacter oxydans glucosidase, Uniparc reference UPI0004A87350, Uniprot reference A0A067Z479.

[0574] SEQ ID NO: 426: Codon optimised DNA encoding 6×His Bifidobacterium catenulatum glucosidase, Uniparc reference UPI00050787A2, Uniprot reference A0A087B8Q8.

[0575] SEQ ID NO: 427: Codon optimised DNA encoding 6×His Bionectria ochroleuca glucosidase, Uniparc reference UPI0005965863, Uniprot reference A0A0B7K538.

[0576] SEQ ID NO: 428: Codon optimised DNA encoding 6×His Parcubacteria sp. glucosidase, Uniparc reference UPI0006377CA4, Uniprot reference A0A0G0GD78.

[0577] SEQ ID NO: 429: Codon optimised DNA encoding 6×His Microbacterium ketosireducens glucosidase, Uniparc reference UPI0006228575, Uniprot reference A0A0M2H276.

[0578] SEQ ID NO: 430: Codon optimised DNA encoding 6×His Roseburia faecis glucosidase, Uniparc reference UPI0006C454B4, Uniprot reference A0A173R3W4.

[0579] SEQ ID NO: 431: Codon optimised DNA encoding 6×His Kwoniella dejecticola glucosidase, Uniparc reference UPI0007F1D695, Uniprot reference A0A1A6A050.

[0580] SEQ ID NO: 432: Codon optimised DNA encoding 6×His uncultured Clostridium sp. glucosidase, Uniparc reference UPI0008232A70, Uniprot reference A0A1C6C862.

[0581] SEQ ID NO: 433: Codon optimised DNA encoding 6×His Cyberlindnera jadinii glucosidase, Uniparc reference UPI000866CB30, Uniprot reference A0A1E4S2F8.

[0582] SEQ ID NO: 434: Codon optimised DNA encoding 6×His Bacteroidetes bacterium glucosidase Uniparc reference UPI0009D3483D, Uniprot reference A0A1V5G4W6.

[0583] SEQ ID NO: 435: Codon optimised DNA encoding 6×His Verrucomicrobia bacterium glucosidase, Uniparc reference UPI0009CC9AEA, Uniprot reference A0A1V5Q4R4.

[0584] SEQ ID NO: 436: Codon optimised DNA encoding 6×His Bacteroidetes bacterium glucosidase Uniparc reference UPI0009C6DE73, Uniprot reference A0A1V6BV25.

[0585] SEQ ID NO: 437: Codon optimised DNA encoding 6×His Lichtheimia ramosa glucosidase, Uniparc reference UPI0004E051A9, Uniprot reference A0A077WUK7.

[0586] SEQ ID NO: 438: Codon optimised DNA encoding 6×His Bifidobacterium mongoliense glucosidase, Uniparc reference UPI0005060F52, Uniprot reference A0A087BWT7.

[0587] SEQ ID NO: 439: Codon optimised DNA encoding 6×His Vibrio ishigakensis glucosidase, Uniparc reference UPI000591CED9, Uniprot reference A0A0B8NZY1.

[0588] SEQ ID NO: 440: Codon optimised DNA encoding 6×His Phaeomoniella chlamydospora glucosidase, Uniparc reference UPI00063B706F, Uniprot reference A0A0G2HEV5.

[0589] SEQ ID NO: 441: Codon optimised DNA encoding 6×His Ardenticatena maritima glucosidase, Uniparc reference UPI0006C04F59, Uniprot reference A0A0M8K5H7.

[0590] SEQ ID NO: 442: Codon optimised DNA encoding 6×His Coprococcus comes glucosidase, Uniparc reference UPI000197E031, Uniprot reference A0A173WPC4.

[0591] SEQ ID NO: 443: Codon optimised DNA encoding 6×His Nocardioides dokdonensis glucosidase, Uniparc reference UPI0007DDBAB3, Uniprot reference A0A1A9GNJ0.

[0592] SEQ ID NO: 444: Codon optimised DNA encoding 6×His uncultured Clostridium sp. glucosidase, Uniparc reference UPI000822F7EB, Uniprot reference A0A1C6EBC1.

[0593] SEQ ID NO: 445: Codon optimised DNA encoding 6×His Acetobacterium wieringae glucosidase, Uniparc reference UPI000878EB40, Uniprot reference A0A1F2PFB4.

[0594] SEQ ID NO: 446: Codon optimised DNA encoding 6×His Tenericutes bacterium glucosidase, Uniparc reference UPI0009D0A2C1, Uniprot reference A0A1V5HNJ3.

[0595] SEQ ID NO: 447: Codon optimised DNA encoding 6×His Tenericutes bacterium glucosidase, Uniparc reference UPI0009CB8A73, Uniprot reference A0A1V5UFI1.

[0596] SEQ ID NO: 448: Codon optimised DNA encoding 6×His uncultured bacterium glucosidase, Uniparc reference UPI0009CC90AF, Uniprot reference A0A1V6CDT2.

[0597] SEQ ID NO: 449: Codon optimised DNA encoding 6×His Parabacteroides distasonis glucosidase, Uniparc reference UPI0004D8E473, Uniprot reference A0A078SYD0.

[0598] SEQ ID NO: 450: Codon optimised DNA encoding 6×His Bifidobacterium psychraerophilum glucosidase, Uniparc reference UPI0005006C6A, Uniprot reference A0A087CJB0.

[0599] SEQ ID NO: 451: Codon optimised DNA encoding 6×His Hebeloma cylindrosporum glucosidase, Uniparc reference UPI00059A3BA8, Uniprot reference A0A0C2YJF2.

[0600] SEQ ID NO: 452: Codon optimised DNA encoding 6×His Brenneria goodwinii glucosidase, Uniparc reference UPI0006579FEA, Uniprot reference A0A0G4JRR8.

[0601] SEQ ID NO: 453: Codon optimised DNA encoding 6×His Aspergillus calidoustus glucosidase, Uniparc reference UPI00073C9E60, Uniprot reference A0A0U5GN20.

[0602] SEQ ID NO: 454: Codon optimised DNA encoding 6×His Bacteroides finegoldii glucosidase, Uniparc reference UPI0006C6E0C3, Uniprot reference A0A174BEZ2.

[0603] SEQ ID NO: 455: Codon optimised DNA encoding 6×His Altererythrobacter dongtanensis glucosidase, Uniparc reference UPI0008153D6D, Uniprot reference A0A1B2A943.

[0604] SEQ ID NO: 456: Codon optimised DNA encoding 6×His uncultured Anaerotruncus sp. glucosidase, Uniparc reference UPI0008206E7B, Uniprot reference A0A1C6FWD9.

[0605] SEQ ID NO: 457: Codon optimised DNA encoding 6×His Candidatus firestone glucosidase, Uniparc reference UPI0008AC0E17, Uniprot reference A0A1F5UJR6.

[0606] SEQ ID NO: 458: Codon optimised DNA encoding 6×His Tenericutes bacterium glucosidase, Uniparc reference UPI0009C6C8E4, Uniprot reference A0A1V5HS22.

[0607] SEQ ID NO: 459: Codon optimised DNA encoding 6×His Lentisphaerae bacterium glucosidase, Uniparc reference UPI0009CEE1AB, Uniprot reference A0A1V5VBL2.

[0608] SEQ ID NO: 460: Codon optimised DNA encoding 6×His Planctomycetes bacterium glucosidase, Uniparc reference UPI0009CAAB0A, Uniprot reference A0A1V6FZ47.

[0609] SEQ ID NO: 461: Codon optimised DNA encoding 6×His Pseudallescheria apiosperma glucosidase, Uniparc reference UPI0004DD62AC, Uniprot reference A0A084G332.

[0610] SEQ ID NO: 462: Codon optimised DNA encoding 6×His Nonlabens sediminis glucosidase, Uniparc reference UPI000507F00A, Uniprot reference A0A090Q4N8.

[0611] SEQ ID NO: 463: Codon optimised DNA encoding 6×His Gynuella sunshinyii glucosidase, Uniparc reference UPI0005CC42CA, Uniprot reference A0A0C5VDU3.

[0612] SEQ ID NO: 464: Codon optimised DNA encoding 6×His Verticillium longisporum glucosidase, Uniparc reference UPI00063E4005, Uniprot reference A0A0G4N9Q7.

[0613] SEQ ID NO: 465: Codon optimised DNA encoding 6×His Cellulomonas sp. glucosidase, Uniparc reference UPI00073C6CD3, Uniprot reference A0A0V8TAB5.

[0614] SEQ ID NO: 466: Codon optimised DNA encoding 6×His Hungatella hathewayi glucosidase, Uniparc reference UPI0006C024BB, Uniprot reference A0A174FBX7.

[0615] SEQ ID NO: 467: Codon optimised DNA encoding 6×His Mesorhizobium sp. glucosidase, Uniparc reference UPI000688E74D, Uniprot reference A0A1C2DG64.

[0616] SEQ ID NO: 468: Codon optimised DNA encoding 6×His Clostridium sp. glucosidase, Uniparc reference UPI000822FAB7, Uniprot reference A0A1C6GRT5.

[0617] SEQ ID NO: 469: Codon optimised DNA encoding 6×His Chlamydiales bacterium glucosidase, Uniparc reference UPI0009284E74, Uniprot reference A0A1M3CSY6.

[0618] SEQ ID NO: 470: Codon optimised DNA encoding 6×His Spirochaetes bacterium glucosidase, Uniparc reference UPI0009CB461D, Uniprot reference A0A1V5HUX3.

[0619] SEQ ID NO: 471: Codon optimised DNA encoding 6×His bacterium glucosidase, Uniparc reference UPI0009C92D6A, Uniprot reference A0A1V5VHB9.

[0620] SEQ ID NO: 472: Codon optimised DNA encoding 6×His Thermotogae bacterium glucosidase, Uniparc reference UPI0009D12B86, Uniprot reference A0A1V6H2W5.

[0621] SEQ ID NO: 473: Codon optimised DNA encoding 6×His Pseudallescheria apiosperma glucosidase, Uniparc reference UPI0004DCBED3, Uniprot reference A0A084GGE2.

[0622] SEQ ID NO: 474: Codon optimised DNA encoding 6×His Algibacter lectus glucosidase, Uniparc reference UPI00050E0BE3, Uniprot reference A0A090VF17.

[0623] SEQ ID NO: 475: Codon optimised DNA encoding 6×His Paxillus involutus glucosidase, Uniparc reference UPI0005B075C8, Uniprot reference A0A0C9TWP5.

[0624] SEQ ID NO: 476: Codon optimised DNA encoding 6×His Verticillium longisporum glucosidase, Uniparc reference UPI00063DF006, Uniprot reference A0A0G4NA55.

[0625] SEQ ID NO: 477: Codon optimised DNA encoding 6×His Mucilaginibacter gotjawali glucosidase, Uniparc reference UPI00076F8EA4, Uniprot reference A0A110B1H1.

[0626] SEQ ID NO: 478: Codon optimised DNA encoding 6×His Bacteroides uniformis glucosidase, Uniparc reference UPI0006C0FFEF, Uniprot reference A0A174IWW4.

[0627] SEQ ID NO: 479: Codon optimised DNA encoding 6×His Coprococcus sp. glucosidase, Uniparc reference UPI0008222B77, Uniprot reference A0A1C5W0N6.

[0628] SEQ ID NO: 480: Codon optimised DNA encoding 6×His Blautia sp. glucosidase, Uniparc reference UPI0006C36823, Uniprot reference A0A1C6K2X5.

[0629] SEQ ID NO: 481: Codon optimised DNA encoding 6×His Cellulomonas sp. glucosidase, Uniparc reference UPI00092B3CD4, Uniprot reference A0A1M3ELH4.

[0630] SEQ ID NO: 482: Codon optimised DNA encoding 6×His bacterium glucosidase, Uniparc reference UPI0009C85428, Uniprot reference A0A1V5J984.

[0631] SEQ ID NO: 483: Codon optimised DNA encoding 6×His Spirochaetes bacterium glucosidase, Uniparc reference UPI0009CA1D5C, Uniprot reference A0A1V5WHQ7.

[0632] SEQ ID NO: 484: Codon optimised DNA encoding 6×His Tenericutes bacterium glucosidase, Uniparc reference UPI0009C7041F, Uniprot reference A0A1V6IJ57.

[0633] SEQ ID NO: 485: Codon optimised DNA encoding 6×His Flavobacterium gilvum glucosidase, Uniparc reference UPI0004E2A41F, Uniprot reference A0A085EG29.

[0634] SEQ ID NO: 486: Codon optimised DNA encoding 6×His Algibacter lectus glucosidase, Uniparc reference UPI0005102470, Uniprot reference A0A090WWZ1.

[0635] SEQ ID NO: 487: Codon optimised DNA encoding 6×His Hydnomerulius pinastri glucosidase, Uniparc reference UPI0005B0F0AE, Uniprot reference A0A0C9WDY0.

[0636] SEQ ID NO: 488: Codon optimised DNA encoding 6×His Nocardia farcinica glucosidase, Uniparc reference UPI00065C2666, Uniprot reference A0A0H5NWN2.

[0637] SEQ ID NO: 489: Codon optimised DNA encoding 6×His Bacteroides cellulosilyticus glucosidase, Uniparc reference UPI000760375B, Uniprot reference A0A125MG18.

[0638] SEQ ID NO: 490: Codon optimised DNA encoding 6×His Fonsecaea erecta glucosidase, Uniparc reference UPI0007DF4250, Uniprot reference A0A178ZBN2.

[0639] SEQ ID NO: 491: Codon optimised DNA encoding 6×His Bacteroides sp. glucosidase, Uniparc reference UPI0008209852, Uniprot reference A0A1C5W9N9.

[0640] SEQ ID NO: 492: Codon optimised DNA encoding 6×His Tannerella forsythia glucosidase, Uniparc reference UPI00086C34C1, Uniprot reference A0A1D3UGH8.

[0641] SEQ ID NO: 493: Codon optimised DNA encoding 6×His Microbacterium esteraromaticum glucosidase, Uniparc reference UPI00097E83BB, Uniprot reference A0A1 R4KI92.

[0642] SEQ ID NO: 494: Codon optimised DNA encoding 6×His bacterium glucosidase, Uniparc reference UPI0009D255E5, Uniprot reference A0A1V5LJK9.

[0643] SEQ ID NO: 495: Codon optimised DNA encoding 6×His Candidatus hydrogenedentes glucosidase, Uniparc reference UPI0009C5A3CF, Uniprot reference A0A1V5Z2L2.

[0644] SEQ ID NO: 496: Codon optimised DNA encoding 6×His Bacteroidetes bacterium glucosidase, Uniparc reference UPI0009C55799, Uniprot reference A0A1V6J4J8.

[0645] SEQ ID NO: 497: Codon optimised DNA encoding 6×His Penicillium solitum glucosidase, Uniparc reference UPI0009D4067F, Uniprot reference A0A1V6RQ41.

[0646] SEQ ID NO: 498: Codon optimised DNA encoding 6×His Weissella soli glucosidase, Uniparc reference UPI0008737AA2, Uniprot reference A0A288Q812.

[0647] SEQ ID NO: 499: Codon optimised DNA encoding 6×His Acetatifactor muris glucosidase, Uniparc reference UPI000CAC57D4, Uniprot reference A0A2K4ZN91.

[0648] SEQ ID NO: 500: Codon optimised DNA encoding 6×His Corynespora cassiicola glucosidase, Uniparc reference UPI000D237A4A, Uniprot reference A0A2T2NYD4.

[0649] SEQ ID NO: 501: Codon optimised DNA encoding 6×His Meira miltonrushii glucosidase, Uniparc reference UPI000D77C91D, Uniprot reference A0A316V6M3.

[0650] SEQ ID NO: 502: Codon optimised DNA encoding 6×His Bacteroides fragilis glucosidase, Uniparc reference UPI00004E1F76, Uniprot reference A0A380YVC7.

[0651] SEQ ID NO: 503: Codon optimised DNA encoding 6×His Malassezia restricta glucosidase, Uniparc reference UPI000F0C30E8, Uniprot reference A0A3G2SB79.

[0652] SEQ ID NO: 504: Codon optimised DNA encoding 6×His Fusarium euwallaceae glucosidase, Uniparc reference UPI000FFFEFB2, Uniprot reference A0A430LYA2.

[0653] SEQ ID NO: 505: Codon optimised DNA encoding 6×His Psathyrella aberdarensis glucosidase, Uniparc reference UPI0010251887, Uniprot reference A0A4Q2E070.

[0654] SEQ ID NO: 506: Codon optimised DNA encoding 6×His Aeromonas hydrophila glucosidase, Uniparc reference UPI0000E69509, Uniprot reference A0KLP6.

[0655] SEQ ID NO: 507: Codon optimised DNA encoding 6×His Saccharopolyspora erythraea glucosidase, Uniparc reference UPI00000B86CB, Uniprot reference A4F7P9.

[0656] SEQ ID NO: 508: Codon optimised DNA encoding 6×His Streptomyces sviceus glucosidase, Uniparc reference UPI000180240E, Uniprot reference B51181.

[0657] SEQ ID NO: 509: Codon optimised DNA encoding 6×His Naematelia encephala glucosidase, Uniparc reference UPI000A250F78, Uniprot reference A0A1Y2AWB7.

[0658] SEQ ID NO: 510: Codon optimised DNA encoding 6×His Hartmannibacter diazotrophicus glucosidase, Uniparc reference UPI000C0221F1, Uniprot reference A0A2C9D612.

[0659] SEQ ID NO: 511: Codon optimised DNA encoding 6×His Pontimonas salivibrio glucosidase, Uniparc reference UPI000CEB5AB1, Uniprot reference A0A2L2BPE2.

[0660] SEQ ID NO: 512: Codon optimised DNA encoding 6×His Cadophora sp. glucosidase, Uniparc reference UPI000D5B9C38, Uniprot reference A0A2V1CH24.

[0661] SEQ ID NO: 513: Codon optimised DNA encoding 6×His Meira miltonrushii glucosidase, Uniparc reference UPI000D779558, Uniprot reference A0A316V8S0.

[0662] SEQ ID NO: 514: Codon optimised DNA encoding 6×His Monilinia fructigena glucosidase, Uniparc reference UPI000DC42E3D, Uniprot reference A0A395IJW4.

[0663] SEQ ID NO: 515: Codon optimised DNA encoding 6×His Hortaea werneckii glucosidase, Uniparc reference UPI000F3E476C, Uniprot reference A0A3M6XGS0.

[0664] SEQ ID NO: 516: Codon optimised DNA encoding 6×His Streptomyces netropsis glucosidase, Uniparc reference UPI00101460D7, Uniprot reference A0A445N7U0.

[0665] SEQ ID NO: 517: Codon optimised DNA encoding 6×His Aureobasidium pullulans glucosidase, Uniparc reference UPI001139C6A8, Uniprot reference A0A4S9IFI0.

[0666] SEQ ID NO: 518: Codon optimised DNA encoding 6×His Aspergillus clavatus glucosidase, Uniparc reference UPI0000EA5CFF, Uniprot reference A1CTN9.

[0667] SEQ ID NO: 519: Codon optimised DNA encoding 6×His Clavibacter michiganensis glucosidase, Uniparc reference UPI0001523037, Uniprot reference A5CT94.

[0668] SEQ ID NO: 520: Codon optimised DNA encoding 6×His Penicillium rubens glucosidase, Uniparc reference UPI0001831CF5, Uniprot reference B6H7R5.

[0669] SEQ ID NO: 521: Codon optimised DNA encoding 6×His Lachnoclostridium sp. glucosidase, Uniparc reference UPI000B365547, Uniprot reference A0A1Y4NTL9.

[0670] SEQ ID NO: 522: Codon optimised DNA encoding 6×His Rhodobacteraceae bacterium glucosidase, Uniparc reference UPI000C09BF88, Uniprot reference A0A2D5IXB9.

[0671] SEQ ID NO: 523: Codon optimised DNA encoding 6×His Bacteroides fragilis glucosidase, Uniparc reference UPI0004B5EEF2, Uniprot reference A0A2M9UUC4.

[0672] SEQ ID NO: 524: Codon optimised DNA encoding 6×His Aspergillus indologenus glucosidase, Uniparc reference UPI000D7FE1 ED, Uniprot reference A0A2V51Y78.

[0673] SEQ ID NO: 525: Codon optimised DNA encoding 6×His Acaromyces ingoldii glucosidase, Uniparc reference UPI000D802B25, Uniprot reference A0A316YR39.

[0674] SEQ ID NO: 526: Codon optimised DNA encoding 6×His Monilinia fructigena glucosidase, Uniparc reference UPI000DC60823, Uniprot reference A0A395J1U5.

[0675] SEQ ID NO: 527: Codon optimised DNA encoding 6×His Paenibacillus xylanexedens glucosidase, Uniparc reference UPI000F52D773, Uniprot reference A0A3N6CA02.

[0676] SEQ ID NO: 528: Codon optimised DNA encoding 6×His Actinomyces howellii glucosidase, Uniparc reference UPI000F6DAAAE, Uniprot reference A0A448HIG0.

[0677] SEQ ID NO: 529: Codon optimised DNA encoding 6×His Friedmanniomyces endolithicus glucosidase, Uniparc reference UPI00113D19DF, Uniprot reference A0A4V5N914.

[0678] SEQ ID NO: 530: Codon optimised DNA encoding 6×His Neosartorya fischeri glucosidase, Uniparc reference UPI0000EA8672, Uniprot reference A1DNS0.

[0679] SEQ ID NO: 531: Codon optimised DNA encoding 6×His Pseudomonas aeruginosa glucosidase, Uniparc reference UPI0000D7314B, Uniprot reference A6V4K6.

[0680] SEQ ID NO: 532: Codon optimised DNA encoding 6×His Talaromyces stipitatus glucosidase, Uniparc reference UPI00018E7266, Uniprot reference B8MF24.

[0681] SEQ ID NO: 533: Codon optimised DNA encoding 6×His Aquimixticola soesokkakensis glucosidase, Uniparc reference UPI000A1A5FD7, Uniprot reference A0A1Y5RVF8.

[0682] SEQ ID NO: 534: Codon optimised DNA encoding 6×His Rhodobacterales bacterium glucosidase, Uniparc reference UPI000C98D37C, Uniprot reference A0A2D9YGV1.

[0683] SEQ ID NO: 535: Codon optimised DNA encoding 6×His Methylorubrum extorquens glucosidase, Uniparc reference UPI0006F9793E, Uniprot reference A0A2N9AS40.

[0684] SEQ ID NO: 536: Codon optimised DNA encoding 6×His Clostridium perfringens glucosidase, Uniparc reference UPI00000CF7C8, Uniprot reference A0A2X2YBP0.

[0685] SEQ ID NO: 537: Codon optimised DNA encoding 6×His Acholeplasmatales bacterium glucosidase, Uniparc reference UPI0008AEBEA3, Uniprot reference A0A348NID6.

[0686] SEQ ID NO: 538: Codon optimised DNA encoding 6×His Bacteroidetes bacterium glucosidase, Uniparc reference UPI000EC3C979, Uniprot reference A0A3B8VHE8.

[0687] SEQ ID NO: 539: Codon optimised DNA encoding 6×His Clostridium carnis glucosidase, Uniparc reference UPI000F637E38, Uniprot reference A0A3P6K8E8.

[0688] SEQ ID NO: 540: Codon optimised DNA encoding 6×His Mycolicibacterium flavescens glucosidase, Uniparc reference UPI000B93B5C9, Uniprot reference A0A448HNB5.

[0689] SEQ ID NO: 541: Codon optimised DNA encoding 6×His Streptococcus gallolyticus glucosidase, Uniparc reference UPI000F6EFA71, Uniprot reference A0A4V6LJ94.

[0690] SEQ ID NO: 542: Codon optimised DNA encoding 6×His Yersinia enterocolitica glucosidase, Uniparc reference UPI0000EB54CC, Uniprot reference A1JNB7.

[0691] SEQ ID NO: 543: Codon optimised DNA encoding 6×His Anaeromyxobacter sp. glucosidase, Uniparc reference UPI0000ED8A80, Uniprot reference A7HFG4.

[0692] SEQ ID NO: 544: Codon optimised DNA encoding 6×His Talaromyces stipitatus glucosidase, Uniparc reference UPI00018E7D70, Uniprot reference B8MK55.

[0693] SEQ ID NO: 545: Codon optimised DNA encoding 6×His Hortaea werneckii glucosidase, Uniparc reference UPI000A2E3FAA, Uniprot reference A0A1Z5SL14.

[0694] SEQ ID NO: 546: Codon optimised DNA encoding 6×His Micavibrio sp. glucosidase, Uniparc reference UPI000C529025, Uniprot reference A0A2E2Q8X2.

[0695] SEQ ID NO: 547: Codon optimised DNA encoding 6×His Acidobacteriia bacterium glucosidase, Uniparc reference UPI0000CE6B996, Uniprot reference A0A2N9MBS0.

[0696] SEQ ID NO: 548: Codon optimised DNA encoding 6×His Corynebacterium jeikeium glucosidase, Uniparc reference UPI000DA3A972, Uniprot reference A0A2X4T570.

[0697] SEQ ID NO: 549: Codon optimised DNA encoding 6×His Clostridiaceae bacterium glucosidase, Uniparc reference UPI000E8D37A1, Uniprot reference A0A353PZH8.

[0698] SEQ ID NO: 550: Codon optimised DNA encoding 6×His Anaerolineaceae bacterium glucosidase, Uniparc reference UPI000748C096, Uniprot reference A0A3B9PA35.

[0699] SEQ ID NO: 551: Codon optimised DNA encoding 6×His Gymnopilus dilepis glucosidase, Uniparc reference UPI000FF41956, Uniprot reference A0A409WSY0.

[0700] SEQ ID NO: 552: Codon optimised DNA encoding 6×His Kocuria rosea glucosidase, Uniparc reference UPI000F7105D4, Uniprot reference A0A448R8N0.

[0701] SEQ ID NO: 553: Codon optimised DNA encoding 6×His Teredinibacter sp. glucosidase, Uniparc reference UPI0011696FAB, Uniprot reference A0A509DWZ3.

[0702] SEQ ID NO: 554: Codon optimised DNA encoding 6×His Aspergillus niger glucosidase, Uniparc reference UPI0000EFB564, Uniprot reference A2QS42.

[0703] SEQ ID NO: 555: Codon optimised DNA encoding 6×His Laccaria bicolor glucosidase, Uniparc reference UPI000164423D, Uniprot reference B0D734.

[0704] SEQ ID NO: 556: Codon optimised DNA encoding 6×His Pedosphaera parvula glucosidase, Uniparc reference UPI00017357F6, Uniprot reference B9XH33.

[0705] SEQ ID NO: 557: Codon optimised DNA encoding 6×His Megamonas hypermegale glucosidase, Uniparc reference UPI00042469F6, Uniprot reference A0A239TGH2.

[0706] SEQ ID NO: 558: Codon optimised DNA encoding 6×His Armillaria gallica glucosidase, Uniparc reference UPI000BC209C3, Uniprot reference A0A2H3E300.

[0707] SEQ ID NO: 559: Codon optimised DNA encoding 6×His Micromonospora sp. glucosidase, Uniparc reference UPI000D2EAE87, Uniprot reference A0A2P8AV03.

[0708] SEQ ID NO: 560: Codon optimised DNA encoding 6×His Klebsiella oxytoca glucosidase, Uniparc reference UPI000DA286EE, Uniprot reference A0A2X5CJC5.

[0709] SEQ ID NO: 561: Codon optimised DNA encoding 6×His Candidatus ozemobacter glucosidase, Uniparc reference UPI000DFAEF6C, Uniprot reference A0A367Z1H8.

[0710] SEQ ID NO: 562: Codon optimised DNA encoding 6×His Coleophoma crateriformis glucosidase, Uniparc reference UPI000E399EE8, Uniprot reference A0A3D8R2C2.

[0711] SEQ ID NO: 563: Codon optimised DNA encoding 6×His Apiotrichum porosum glucosidase, Uniparc reference UPI000FBC01E3, Uniprot reference A0A427XHS2.

[0712] SEQ ID NO: 564: Codon optimised DNA encoding 6×His Acholeplasma hippikon glucosidase, Uniparc reference UPI00068E4E50, Uniprot reference A0A449BJ27.

[0713] SEQ ID NO: 565: Codon optimised DNA encoding 6×His Streptomyces spectabilis glucosidase, Uniparc reference UPI001185F074, Uniprot reference A0A516RGT1.

[0714] SEQ ID NO: 566: Codon optimised DNA encoding 6×His Aspergillus niger glucosidase, Uniparc reference UPI0000EFCED2, Uniprot reference A2R8G2.

[0715] SEQ ID NO: 567: Codon optimised DNA encoding 6×His Xanthomonas campestris glucosidase, Uniparc reference UPI00000D8BFA, Uniprot reference B0RYA0.

[0716] SEQ ID NO: 568: Codon optimised DNA encoding 6×His Lactobacillus paracasei glucosidase, Uniparc reference UPI00019C9CD7, Uniprot reference C2FDL2.

[0717] SEQ ID NO: 569: Codon optimised DNA encoding 6×His Bifiguratus adelaidae glucosidase, Uniparc reference UPI000BC490A3, Uniprot reference A0A261XUH4.

[0718] SEQ ID NO: 570: Codon optimised DNA encoding 6×His bacterium glucosidase, Uniparc reference UPI000CC31AE2, Uniprot reference A0A2H5YYA1.

[0719] SEQ ID NO: 571: Codon optimised DNA encoding 6×His Actinomadura parvosata glucosidase, Uniparc reference UPI000D26C9C6, Uniprot reference A0A2P91Y35.

[0720] SEQ ID NO: 572: Codon optimised DNA encoding 6×His Melissococcus plutonius glucosidase, Uniparc reference UPI00024F22B8, Uniprot reference A0A2Z5Y4P3.

[0721] SEQ ID NO: 573: Codon optimised DNA encoding 6×His Enterococcus durans glucosidase, Uniparc reference UPI000E020871, Uniprot reference A0A377KJS3.

[0722] SEQ ID NO: 574: Codon optimised DNA encoding 6×His Malassezia restricta glucosidase, Uniparc reference UPI000DD17A7D, Uniprot reference A0A3G2S2J6.

[0723] SEQ ID NO: 575: Codon optimised DNA encoding 6×His Apiotrichum porosum glucosidase, Uniparc reference UPI000FA2DE87, Uniprot reference A0A427XZQ0.

[0724] SEQ ID NO: 576: Codon optimised DNA encoding 6×His Streptomonospora sp. glucosidase, Uniparc reference UPI0010355193, Uniprot reference A0A4P6PWP5.

[0725] SEQ ID NO: 577: Codon optimised DNA encoding 6×His Lactobacillus gasseri glucosidase, Uniparc reference UPI00119640A1, Uniprot reference A0A558LH47.

[0726] SEQ ID NO: 578: Codon optimised DNA encoding 6×His Aspergillus niger glucosidase, Uniparc reference UPI0000EFD0AA, Uniprot reference A2RAJ1.

[0727] SEQ ID NO: 579: Codon optimised DNA encoding 6×His Neosartorya fumigata glucosidase, Uniparc reference UPI000170BF91, Uniprot reference B0XXG1.

[0728] SEQ ID NO: 580: Codon optimised DNA encoding 6×His Eubacterium eligens glucosidase, Uniparc reference UPI0001A5B234, Uniprot reference C4Z6T5.

[0729] SEQ ID NO: 581: Codon optimised DNA encoding 6×His Bifiguratus adelaidae glucosidase, Uniparc reference UPI000BC64A75, Uniprot reference A0A261XVM8.

[0730] SEQ ID NO: 582: Codon optimised DNA encoding 6×His bacterium glucosidase, Uniparc reference UPI000CAA854E, Uniprot reference A0A2H6EX57.

[0731] SEQ ID NO: 583: Codon optimised DNA encoding 6×His Corynespora cassiicola glucosidase, Uniparc reference UPI000D24EC2C, Uniprot reference A0A2T2N5H9.

[0732] SEQ ID NO: 584: Codon optimised DNA encoding 6×His Pseudomicrostroma glucosiphilum glucosidase, Uniparc reference UPI000D77A0B7, Uniprot reference A0A316UK83.

[0733] SEQ ID NO: 585: Codon optimised DNA encoding 6×His Staphylococcus saprophyticus glucosidase, Uniparc reference UPI0002DAAFCC, Uniprot reference A0A380HD31.

[0734] SEQ ID NO: 586: Codon optimised DNA encoding 6×His Malassezia restricta glucosidase, Uniparc reference UPI000F0C6B8E, Uniprot reference A0A3G2S932.

[0735] SEQ ID NO: 587: Codon optimised DNA encoding 6×His Saitozyma podzolica glucosidase, Uniparc reference UPI000FBA70E2, Uniprot reference A0A427YCL1.

[0736] SEQ ID NO: 588: Codon optimised DNA encoding 6×His Tremella mesenterica glucosidase, Uniparc reference UPI00102822B4, Uniprot reference A0A4Q1BRC6.

[0737] SEQ ID NO: 589: Codon optimised DNA encoding 6×His Arthrobacter sp. glucosidase, Uniparc reference UPI0000527506, Uniprot reference A0JZ86.

[0738] SEQ ID NO: 590: Codon optimised DNA encoding 6×His Scheffersomyces stipitis glucosidase, Uniparc reference UPI000157388C, Uniprot reference A3LRB0.

[0739] SEQ ID NO: 591: Codon optimised DNA encoding 6×His Leptothrix cholodnii glucosidase, Uniparc reference UPI0001712E02, Uniprot reference B1XZK8.

[0740] SEQ ID NO: 592: Codon optimised DNA encoding 6×His Thauera sp. glucosidase, Uniparc reference UPI000166883C, Uniprot reference C4ZLL7.

[0741] SEQ ID NO: 593: Codon optimised DNA encoding 6×His Kosmotoga olearia glucosidase, Uniparc reference UPI00018494AB, Uniprot reference C5CDW5.

[0742] SEQ ID NO: 594: Codon optimised DNA encoding 6×His Roseburia intestinalis glucosidase, Uniparc reference UPI0001CD671F, Uniprot reference D4L3Y2.

[0743] SEQ ID NO: 595: Codon optimised DNA encoding 6×His Streptococcus equinus glucosidase, Uniparc reference UPI0001E0DC00, Uniprot reference E0PDF8.

[0744] SEQ ID NO: 596: Codon optimised DNA encoding 6×His Streptococcus cristatus glucosidase, Uniparc reference UPI0001F80C4B, Uniprot reference E8JUK5.

[0745] SEQ ID NO: 597: Codon optimised DNA encoding 6×His Cellulosilyticum lentocellum glucosidase, Uniparc reference UPI0001D2DBBC, Uniprot reference F2JLH3.

[0746] SEQ ID NO: 598: Codon optimised DNA encoding 6×His Streptococcus gallolyticus glucosidase, Uniparc reference UPI0001C48657, Uniprot reference F5WYI5.

[0747] SEQ ID NO: 599: Codon optimised DNA encoding 6×His Ketogulonicigenium vulgare glucosidase, Uniparc reference UPI00021D443A, Uniprot reference F9Y8Z7.

[0748] SEQ ID NO: 600: Codon optimised DNA encoding 6×His Spathaspora passalidarum glucosidase, Uniparc reference UPI000228255D, Uniprot reference G3AGX1.

[0749] SEQ ID NO: 601: Codon optimised DNA encoding 6×His Niastella koreensis glucosidase, Uniparc reference UPI00023F6F5F, Uniprot reference G8T9J3.

[0750] SEQ ID NO: 602: Codon optimised DNA encoding 6×His Cellvibrio sp. glucosidase, Uniparc reference UPI000260108C, Uniprot reference I3IDC0.

[0751] SEQ ID NO: 603: Codon optimised DNA encoding 6×His Flavobacterium sp. glucosidase, Uniparc reference UPI000272D1E0, Uniprot reference J1ACA0.

[0752] SEQ ID NO: 604: Codon optimised DNA encoding 6×His Macrophomina phaseolina glucosidase, Uniparc reference UPI00028E7FE1, Uniprot reference K2S5D3.

[0753] SEQ ID NO: 605: Codon optimised DNA encoding 6×His Kosmotoga olearia glucosidase, Uniparc reference UPI00018483A2, Uniprot reference C5CHI5.

[0754] SEQ ID NO: 606: Codon optimised DNA encoding 6×His Blautia obeum glucosidase, Uniparc reference UPI0001CD5918, Uniprot reference D4LRF6.

[0755] SEQ ID NO: 607: Codon optimised DNA encoding 6×His Bifidobacterium dentium glucosidase, Uniparc reference UPI0001E18CDA, Uniprot reference E0Q541.

[0756] SEQ ID NO: 608: Codon optimised DNA encoding 6×His Anaerolinea thermophila glucosidase, Uniparc reference UPI0001F55F8F, Uniprot reference E8N5R8.

[0757] SEQ ID NO: 609: Codon optimised DNA encoding 6×His Coriobacterium glomerans glucosidase, Uniparc reference UPI0002050DA2, Uniprot reference F2N7E4.

[0758] SEQ ID NO: 610: Codon optimised DNA encoding 6×His Microlunatus phosphovorus glucosidase, Uniparc reference UPI000210C886, Uniprot reference F5XJQ3.

[0759] SEQ ID NO: 611: Codon optimised DNA encoding 6×His Streptomyces sp. glucosidase, Uniparc reference UPI0001C18877, Uniprot reference G0Q1U8.

[0760] SEQ ID NO: 612: Codon optimised DNA encoding 6×His Spathaspora passalidarum glucosidase, Uniparc reference UPI0002282B61, Uniprot reference G3AIV6.

[0761] SEQ ID NO: 613: Codon optimised DNA encoding 6×His Glarea lozoyensis glucosidase, Uniparc reference UPI0002402F4A, Uniprot reference H0ET34.

[0762] SEQ ID NO: 614: Codon optimised DNA encoding 6×His Glaciozyma antarctica glucosidase, Uniparc reference UPI0002633B36, Uniprot reference I3UJK0.

[0763] SEQ ID NO: 615: Codon optimised DNA encoding 6×His Arthrobacter sp. glucosidase, Uniparc reference UPI00027DFD35, Uniprot reference J7LN00.

[0764] SEQ ID NO: 616: Codon optimised DNA encoding 6×His Fusarium pseudograminearum glucosidase, Uniparc reference UPI00028D698C, Uniprot reference K3VMA9.

[0765] SEQ ID NO: 617: Codon optimised DNA encoding 6×His Hypocrea rufa glucosidase, Uniparc reference UPI000006AA61, Uniprot reference C6GGC9.

[0766] SEQ ID NO: 618: Codon optimised DNA encoding 6×His Ruminococcus torques glucosidase, Uniparc reference UPI0001CDA964, Uniprot reference D4M6W6.

[0767] SEQ ID NO: 619: Codon optimised DNA encoding 6×His Bifidobacterium dentium glucosidase, Uniparc reference UPI0001E17370, Uniprot reference E0Q9Z7.

[0768] SEQ ID NO: 620: Codon optimised DNA encoding 6×His Bacteroides salanitronis glucosidase, Uniparc reference UPI0001FC71F6, Uniprot reference F0R2D7.

[0769] SEQ ID NO: 621: Codon optimised DNA encoding 6×His Bacteroides coprosuis glucosidase, Uniparc reference UPI00020E6D19, Uniprot reference F3ZQ40.

[0770] SEQ ID NO: 622: Codon optimised DNA encoding 6×His Marinomonas posidonica glucosidase, Uniparc reference UPI00020D4CC6, Uniprot reference F6CWF6.

[0771] SEQ ID NO: 623: Codon optimised DNA encoding 6×His Chaetomium thermophilum glucosidase, Uniparc reference UPI000227E8ED, Uniprot reference G0 SE64.

[0772] SEQ ID NO: 624: Codon optimised DNA encoding 6×His Tetragenococcus halophilus glucosidase, Uniparc reference UPI00022B9A15, Uniprot reference G4L5K1.

[0773] SEQ ID NO: 625: Codon optimised DNA encoding 6×His Paenibacillus sp. glucosidase, Uniparc reference UPI00024F0867, Uniprot reference H6CCZ8.

[0774] SEQ ID NO: 626: Codon optimised DNA encoding 6×His Turneriella parva glucosidase, Uniparc reference UPI000265AA4B, Uniprot reference 14B8U7.

[0775] SEQ ID NO: 627: Codon optimised DNA encoding 6×His Arthrobacter sp. glucosidase, Uniparc reference UPI00027DFDB2, Uniprot reference J7LQK9.

[0776] SEQ ID NO: 628: Codon optimised DNA encoding 6×His Agaricus bisporus glucosidase, Uniparc reference UPI00029074B1, Uniprot reference K5W7V1.

[0777] SEQ ID NO: 629: Codon optimised DNA encoding 6×His Nectria haematococca glucosidase, Uniparc reference UPI0001B67634, Uniprot reference C7YIP3.

[0778] SEQ ID NO: 630: Codon optimised DNA encoding 6×His Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0001A25287, Uniprot reference D4VSZ0.

[0779] SEQ ID NO: 631: Codon optimised DNA encoding 6×His Sediminispirochaeta smaragdinae glucosidase, Uniparc reference UPI0001DD9790, Uniprot reference E1R331.

[0780] SEQ ID NO: 632: Codon optimised DNA encoding 6×His Deinococcus proteolyticus glucosidase, Uniparc reference UPI0001FC42E9, Uniprot reference F0RPV2.

[0781] SEQ ID NO: 633: Codon optimised DNA encoding 6×His Sphingobacterium sp. glucosidase, Uniparc reference UPI0002033AOA, Uniprot reference F4C226.

[0782] SEQ ID NO: 634: Codon optimised DNA encoding 6×His Sphingobium chlorophenolicum glucosidase, Uniparc reference UPI0001E54133, Uniprot reference F6ET40.

[0783] SEQ ID NO: 635: Codon optimised DNA encoding 6×His Caloramator australicus glucosidase, Uniparc reference UPI00021CACC4, Uniprot reference G0V3V5.

[0784] SEQ ID NO: 636: Codon optimised DNA encoding 6×His Commensalibacter intestini glucosidase, Uniparc reference UPI000230E3BF, Uniprot reference G6F370.

[0785] SEQ ID NO: 637: Codon optimised DNA encoding 6×His Paenibacillus sp. glucosidase, Uniparc reference UPI00024F07AC, Uniprot reference H6CIT2.

[0786] SEQ ID NO: 638: Codon optimised DNA encoding 6×His Nitrolancea hollandica glucosidase, Uniparc reference UPI0002638AF3, Uniprot reference I4EIA9.

[0787] SEQ ID NO: 639: Codon optimised DNA encoding 6×His Cryptococcus neoformans glucosidase, Uniparc reference UPI000392C3ED, Uniprot reference J9VVK7.

[0788] SEQ ID NO: 640: Codon optimised DNA encoding 6×His Acidipropionibacterium acidipropionici glucosidase, Uniparc reference UPI0002988588, Uniprot reference K7S596.

[0789] SEQ ID NO: 641: Codon optimised DNA encoding 6×His Prevotella sp. glucosidase, Uniparc reference UPI0001B93465, Uniprot reference C9PT75.

[0790] SEQ ID NO: 642: Codon optimised DNA encoding 6×His Rhodobacter capsulatus glucosidase, Uniparc reference UPI0001D08095, Uniprot reference D5ALU0.

[0791] SEQ ID NO: 643: Codon optimised DNA encoding 6×His Stigmatella aurantiaca glucosidase, Uniparc reference UPI0001E74370, Uniprot reference E3FJ05.

[0792] SEQ ID NO: 644: Codon optimised DNA encoding 6×His Sphaerochaeta globosa glucosidase, Uniparc reference UPI0002010060, Uniprot reference F0RVK3.

[0793] SEQ ID NO: 645: Codon optimised DNA encoding 6×His Sphaerochaeta coccoides glucosidase, Uniparc reference UPI000207D78C, Uniprot reference F4GH96.

[0794] SEQ ID NO: 646: Codon optimised DNA encoding 6×His Novosphingobium sp. glucosidase, Uniparc reference UPI00020EFBCB, Uniprot reference F6ICQ5.

[0795] SEQ ID NO: 647: Codon optimised DNA encoding 6×His Arthrobotrys oligospora glucosidase, Uniparc reference UPI000225331F, Uniprot reference G1XH86.

[0796] SEQ ID NO: 648: Codon optimised DNA encoding 6×His Lactococcus lactis glucosidase, Uniparc reference UPI00000C6907, Uniprot reference G6FFS4.

[0797] SEQ ID NO: 649: Codon optimised DNA encoding 6×His Phaeospirillum molischianum glucosidase, Uniparc reference UPI000255313A, Uniprot reference H8FXH7.

[0798] SEQ ID NO: 650: Codon optimised DNA encoding 6×His Modestobacter marinus glucosidase, Uniparc reference UPI0002609C68, Uniprot reference I4EW72.

[0799] SEQ ID NO: 651: Codon optimised DNA encoding 6×His Saccharothrix espanaensis glucosidase, Uniparc reference UPI00028AC337, Uniprot reference K0K125.

[0800] SEQ ID NO: 652: Codon optimised DNA encoding 6×His Cronobacter sakazakii glucosidase, Uniparc reference UPI00029BA293, Uniprot reference K8DAK0.

[0801] SEQ ID NO: 653: Codon optimised DNA encoding 6×His Verticillium alfalfae glucosidase, Uniparc reference UPI0001BBDF1E, Uniprot reference C9SVX1.

[0802] SEQ ID NO: 654: Codon optimised DNA encoding 6×His Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0001CCFD36, Uniprot reference D6CY10.

[0803] SEQ ID NO: 655: Codon optimised DNA encoding 6×His Leadbetterella byssophila glucosidase, Uniparc reference UPI0001EBD98A, Uniprot reference E4RUR0.

[0804] SEQ ID NO: 656: Codon optimised DNA encoding 6×His Sphaerochaeta globosa glucosidase, Uniparc reference UPI00020102A5, Uniprot reference F0RYB6.

[0805] SEQ ID NO: 657: Codon optimised DNA encoding 6×His Sphaerochaeta coccoides glucosidase, Uniparc reference UPI000207D6A0, Uniprot reference F4GLH6.

[0806] SEQ ID NO: 658: Codon optimised DNA encoding 6×His Haloplasma contractile glucosidase, Uniparc reference UPI000212252C, Uniprot reference F7Q0Y2.

[0807] SEQ ID NO: 659: Codon optimised DNA encoding 6×His Nitrospirillum amazonense glucosidase, Uniparc reference UPI0002265447, Uniprot reference G1Y4C7.

[0808] SEQ ID NO: 660: Codon optimised DNA encoding 6×His Azospirillum brasilense glucosidase, Uniparc reference UPI00023420B8, Uniprot reference G8AWD9.

[0809] SEQ ID NO: 661: Codon optimised DNA encoding 6×His Phaeospirillum molischianum glucosidase, Uniparc reference UPI000255314F, Uniprot reference H8FXJ8.

[0810] SEQ ID NO: 662: Codon optimised DNA encoding 6×His Modestobacter marinus glucosidase, Uniparc reference UPI000260A2FA, Uniprot reference I4EYD5.

[0811] SEQ ID NO: 663: Codon optimised DNA encoding 6×His Wickerhamomyces ciferrii glucosidase, Uniparc reference UPI000283EB8D, Uniprot reference K0KVJ2.

[0812] SEQ ID NO: 664: Codon optimised DNA encoding 6×His Gloeocapsa sp. glucosidase, Uniparc reference UPI0002A5D085, Uniprot reference K9XKL8.

[0813] SEQ ID NO: 665: Codon optimised DNA encoding 6×His Sphaerobacter thermophilus glucosidase, Uniparc reference UPI0001A3BCB6, Uniprot reference D1C7U8.

[0814] SEQ ID NO: 666: Codon optimised DNA encoding 6×His Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0001CCEF1F, Uniprot reference D6D4V2.

[0815] SEQ ID NO: 667: Codon optimised DNA encoding 6×His Prevotella buccae glucosidase, Uniparc reference UPI0001F149E8, Uniprot reference E6K4W5.

[0816] SEQ ID NO: 668: Codon optimised DNA encoding 6×His Grosmannia clavigera glucosidase, Uniparc reference UPI0001FF1101, Uniprot reference F0XBR0.

[0817] SEQ ID NO: 669: Codon optimised DNA encoding 6×His Melampsora larici-populina glucosidase, Uniparc reference UPI00020F9774, Uniprot reference F4R4W2.

[0818] SEQ ID NO: 670: Codon optimised DNA encoding 6×His Prevotella multisaccharivorax glucosidase, Uniparc reference UPI0002138E9C, Uniprot reference F8N7G1.

[0819] SEQ ID NO: 671: Codon optimised DNA encoding 6×His Streptomyces zinciresistens glucosidase, Uniparc reference UPI0002255A63, Uniprot reference G2G8K4.

[0820] SEQ ID NO: 672: Codon optimised DNA encoding 6×His Granulicella mallensis glucosidase, Uniparc reference UPI0001D9FC40, Uniprot reference G8NY42.

[0821] SEQ ID NO: 673: Codon optimised DNA encoding 6×His Gibberella zeae glucosidase, Uniparc reference UPI00021F1FEE, Uniprot reference I1RH94.

[0822] SEQ ID NO: 674: Codon optimised DNA encoding 6×His Modestobacter marinus glucosidase, Uniparc reference UPI00026090A2, Uniprot reference I4EYK6.

[0823] SEQ ID NO: 675: Codon optimised DNA encoding 6×His Lactobacillus equicursoris glucosidase, Uniparc reference UPI0002869F55, Uniprot reference K0NRS8.

[0824] SEQ ID NO: 676: Codon optimised DNA encoding 6×His Colletotrichum fructicola glucosidase, Uniparc reference UPI0002A93280, Uniprot reference L2F9W0.

[0825] SEQ ID NO: 677: Codon optimised DNA encoding 6×His Streptosporangium roseum glucosidase, Uniparc reference UPI0001 BF8AF6, Uniprot reference D2B261.

[0826] SEQ ID NO: 678: Codon optimised DNA encoding 6×His Listeria grayi glucosidase, Uniparc reference UPI00019F252A, Uniprot reference D7UX19.

[0827] SEQ ID NO: 679: Codon optimised DNA encoding 6×His Enterococcus italicus glucosidase, Uniparc reference UPI0001F11EFF, Uniprot reference E6LF07.

[0828] SEQ ID NO: 680: Codon optimised DNA encoding 6×His Fluviicola taffensis glucosidase, Uniparc reference UPI000203D9EA, Uniprot reference F211T6.

[0829] SEQ ID NO: 681: Codon optimised DNA encoding 6×His Shigella flexneri glucosidase, Uniparc reference UPI00020CA641, Uniprot reference F5N4W9.

[0830] SEQ ID NO: 682: Codon optimised DNA encoding 6×His Actinomyces sp. glucosidase, Uniparc reference UPI0002189080, Uniprot reference F9EFW5.

[0831] SEQ ID NO: 683: Codon optimised DNA encoding 6×His Verticillium dahliae glucosidase, Uniparc reference UPI00022EBE5A, Uniprot reference G2X5V6.

[0832] SEQ ID NO: 684: Codon optimised DNA encoding 6×His Actinoplanes sp. glucosidase, Uniparc reference UPI00023ED5A8, Uniprot reference G8S0M0.

[0833] SEQ ID NO: 685: Codon optimised DNA encoding 6×His Gibberella zeae glucosidase, Uniparc reference UPI000023EA29, Uniprot reference 11S320.

[0834] SEQ ID NO: 686: Codon optimised DNA encoding 6×His Auricularia subglabra glucosidase, Uniparc reference UPI00027CE685, Uniprot reference J0WV06.

[0835] SEQ ID NO: 687: Codon optimised DNA encoding 6×His Nitratireductor indicus glucosidase, Uniparc reference UPI00028E86D6, Uniprot reference K2N268.

[0836] SEQ ID NO: 688: Codon optimised DNA encoding 6×His Thermoclostridium stercorarium glucosidase, Uniparc reference UPI0002AD999E, Uniprot reference L7VH66.

[0837] SEQ ID NO: 689: Codon optimised DNA encoding 6×His Geobacillus sp. glucosidase, Uniparc reference UPI0002AF2DE3, Uniprot reference L8A1S2.

[0838] SEQ ID NO: 690: Codon optimised DNA encoding 6×His uncultured bacterium glucosidase, Uniparc reference UPI000327C3CC, Uniprot reference M9ZC55.

[0839] SEQ ID NO: 691: Codon optimised DNA encoding 6×His Burkholderia ambifaria glucosidase, Uniparc reference UPI000059ACE1, Uniprot reference Q0BAK2.

[0840] SEQ ID NO: 692: Codon optimised DNA encoding 6×His Aspergillus oryzae glucosidase, Uniparc reference UPI0000676B8D, Uniprot reference Q2UIR4.

[0841] SEQ ID NO: 693: Codon optimised DNA encoding 6×His Yersinia pseudotuberculosis glucosidase, Uniparc reference UPI00004269B5, Uniprot reference Q66DJ0.

[0842] SEQ ID NO: 694: Codon optimised DNA encoding 6×His Wallemia ichthyophaga glucosidase, Uniparc reference UPI000331CC98, Uniprot reference R9AF64.

[0843] SEQ ID NO: 695: Codon optimised DNA encoding 6×His Glarea lozoyensis glucosidase, Uniparc reference UPI0003522404, Uniprot reference S3DIM6.

[0844] SEQ ID NO: 696: Codon optimised DNA encoding 6×His Moniliophthora roreri glucosidase, Uniparc reference UPI0003BF753A, Uniprot reference V2YU31.

[0845] SEQ ID NO: 697: Codon optimised DNA encoding 6×His Zhouia amylolytica glucosidase, Uniparc reference UPI0003DBCB4F, Uniprot reference W2ULL3.

[0846] SEQ ID NO: 698: Codon optimised DNA encoding 6×His Fusarium oxysporum glucosidase, Uniparc reference UPI0003F362FF, Uniprot reference W9NQQ3.

[0847] SEQ ID NO: 699: Codon optimised DNA encoding 6×His Flavobacterium johnsoniae glucosidase, Uniparc reference UPI00006EA168, Uniprot reference A5FAA5.

[0848] SEQ ID NO: 700: Codon optimised DNA encoding 6×His Oryza sativa glucosidase, Uniparc reference UPI00002394F5, Uniprot reference Q75193.

[0849] SEQ ID NO: 701: Codon optimised DNA encoding 6×His Clostridium saccharoperbutylacetonicum glucosidase, Uniparc reference UPI0002B65681, Uniprot reference M1MCD1.

[0850] SEQ ID NO: 702: Codon optimised DNA encoding 6×His Rhodococcus sp. glucosidase, Uniparc reference UPI0002D21DB1, Uniprot reference N1MBN6.

[0851] SEQ ID NO: 703: Codon optimised DNA encoding 6×His Burkholderia ambifaria glucosidase, Uniparc reference UPI000059C0CD, Uniprot reference Q0BCV8.

[0852] SEQ ID NO: 704: Codon optimised DNA encoding 6×His Xanthomonas campestris glucosidase, Uniparc reference UPI00005CE9E7, Uniprot reference Q3BVH7.

[0853] SEQ ID NO: 705: Codon optimised DNA encoding 6×His Caulobacter vibrioides glucosidase, Uniparc reference UPI00000C7604, Uniprot reference Q9A6F8.

[0854] SEQ ID NO: 706: Codon optimised DNA encoding 6×His Arcticibacter svalbardensis glucosidase, Uniparc reference UPI000338772A, Uniprot reference R9GRA7.

[0855] SEQ ID NO: 707: Codon optimised DNA encoding 6×His Winogradskyella psychrotolerans glucosidase, Uniparc reference UPI00035A4604, Uniprot reference S7VQ28.

[0856] SEQ ID NO: 708: Codon optimised DNA encoding 6×His Methyloglobulus morosus glucosidase, Uniparc reference UPI0003C4ED6A, Uniprot reference V5DXT8.

[0857] SEQ ID NO: 709: Codon optimised DNA encoding 6×His Pestalotiopsis fici glucosidase, Uniparc reference UPI0003E05C04, Uniprot reference W3WV37.

[0858] SEQ ID NO: 710: Codon optimised DNA encoding 6×His Capronia coronata glucosidase, Uniparc reference UPI000434E698, Uniprot reference W9YNR7.

[0859] SEQ ID NO: 711: Codon optimised DNA encoding 6×His Aspergillus aculeatus glucosidase, Uniparc reference UPI00001268FD, Uniprot reference P48825.

[0860] SEQ ID NO: 712: Codon optimised DNA encoding 6×His Thermotoga neapolitana glucosidase, Uniparc reference UPI0000DD5996, Uniprot reference Q0GC07.

[0861] SEQ ID NO: 713: Codon optimised DNA encoding 6×His Clostridium saccharoperbutylacetonicum glucosidase, Uniparc reference UPI0002B666E8, Uniprot reference M1MJF4.

[0862] SEQ ID NO: 714: Codon optimised DNA encoding 6×His Schizosaccharomyces pombe glucosidase, Uniparc reference UPI000006B001, Uniprot reference O74799.

[0863] SEQ ID NO: 715: Codon optimised DNA encoding 6×His Phaeosphaeria nodorum glucosidase Uniparc reference UPI000161BD2B, Uniprot reference Q0TXF6.

[0864] SEQ ID NO: 716: Codon optimised DNA encoding 6×His Xylella fastidiosa Dixon glucosidase, Uniparc reference UPI00003806C8, Uniprot reference Q3RGJ3.

[0865] SEQ ID NO: 717: Codon optimised DNA encoding 6×His Schizosaccharomyces pombe glucosidase, Uniparc reference UPI000006A330, Uniprot reference Q9P6J6.

[0866] SEQ ID NO: 718: Codon optimised DNA encoding 6×His Arcticibacter svalbardensis glucosidase, Uniparc reference UPI000337E887, Uniprot reference R9GWD6.

[0867] SEQ ID NO: 719: Codon optimised DNA encoding 6×His Colletotrichum gloeosporioides glucosidase, Uniparc reference UPI0003885717, Uniprot reference T0KJI7.

[0868] SEQ ID NO: 720: Codon optimised DNA encoding 6×His uncultured bacterium glucosidase, Uniparc reference UPI0003C9E340, Uniprot reference V5R1E8.

[0869] SEQ ID NO: 721: Codon optimised DNA encoding 6×His Xanthomonas arboricola glucosidase, Uniparc reference UPI0003E06A61, Uniprot reference W4S715.

[0870] SEQ ID NO: 722: Codon optimised DNA encoding 6×His Fusarium oxysporum glucosidase, Uniparc reference UPI00021EC697, Uniprot reference X0A8X8.

[0871] SEQ ID NO: 723: Codon optimised DNA encoding 6×His Paenibacillus polymyxa glucosidase, Uniparc reference UPI00001108D0, Uniprot reference P22073.

[0872] SEQ ID NO: 724: Codon optimised DNA encoding 6×His Kluyveromyces marxianus glucosidase, Uniparc reference UPI0001BE5ADA, Uniprot reference D1GCC6.

[0873] SEQ ID NO: 725: Codon optimised DNA encoding 6×His Ilumatobacter coccineus glucosidase, Uniparc reference UPI0002C04A25, Uniprot reference M5A594.

[0874] SEQ ID NO: 726: Codon optimised DNA encoding 6×His Agrobacterium sp. glucosidase, Uniparc reference UPI0000126912, Uniprot reference P12614.

[0875] SEQ ID NO: 727: Codon optimised DNA encoding 6×His Cytophaga hutchinsonii glucosidase, Uniparc reference UPI000038ECF7, Uniprot reference Q11P53.

[0876] SEQ ID NO: 728: Codon optimised DNA encoding 6×His Thermobifida fusca glucosidase, Uniparc reference UPI00003C5CA4, Uniprot reference Q47PF5.

[0877] SEQ ID NO: 729: Codon optimised DNA encoding 6×His Botryotinia fuckeliana glucosidase, Uniparc reference UPI0000069E5E, Uniprot reference Q9UVJ6.

[0878] SEQ ID NO: 730: Codon optimised DNA encoding 6×His Agarivorans albus glucosidase, Uniparc reference UPI00033994D2, Uniprot reference R9PTK2.

[0879] SEQ ID NO: 731: Codon optimised DNA encoding 6×His Enterococcus sp. glucosidase, Uniparc reference UPI00038B7639, Uniprot reference T0UDU2.

[0880] SEQ ID NO: 732: Codon optimised DNA encoding 6×His Salinispira pacifica glucosidase, Uniparc reference UPI0003D8A7DC, Uniprot reference V5WKT4.

[0881] SEQ ID NO: 733: Codon optimised DNA encoding 6×His Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0003ECF15E, Uniprot reference W6P696.

[0882] SEQ ID NO: 734: Codon optimised DNA encoding 6×His Fusarium oxysporum glucosidase, Uniparc reference UPI000430029A, Uniprot reference X0AB46.

[0883] SEQ ID NO: 735: Codon optimised DNA encoding 6×His Thermotoga maritima glucosidase, Uniparc reference UPI0000126906, Uniprot reference Q08638B.

[0884] SEQ ID NO: 736: Codon optimised DNA encoding 6×His Neotermes koshunensis glucosidase Uniparc reference UPI0000083EA8, Uniprot reference Q8T0W7.

[0885] SEQ ID NO: 737: Codon optimised DNA encoding 6×His Thanatephorus cucumeris glucosidase, Uniparc reference UPI0002BF34B3, Uniprot reference M5CHG9.

[0886] SEQ ID NO: 738: Codon optimised DNA encoding 6×His Hungateiclostridium thermocellum glucosidase, Uniparc reference UPI000053581 D, Uniprot reference P14002.

[0887] SEQ ID NO: 739: Codon optimised DNA encoding 6×His Koribacter versatilis glucosidase, Uniparc reference UPI0000D76A70, Uniprot reference Q1IJ89.

[0888] SEQ ID NO: 740: Codon optimised DNA encoding 6×His Neosartorya fumigata glucosidase, Uniparc reference UPI00005203D8, Uniprot reference Q4WLX5.

[0889] SEQ ID NO: 741: Codon optimised DNA encoding 6×His Amycolatopsis vancoresmycina glucosidase, Uniparc reference UPI00032DB8C1, Uniprot reference R1FKF0.

[0890] SEQ ID NO: 742: Codon optimised DNA encoding 6×His Gibberella fujikuroi glucosidase, Uniparc reference UPI0003519C76, Uniprot reference S0EKU1.

[0891] SEQ ID NO: 743: Codon optimised DNA encoding 6×His Bifidobacterium longum glucosidase, Uniparc reference UPI000390F839, Uniprot reference T212H5.

[0892] SEQ ID NO: 744: Codon optimised DNA encoding 6×His uncultured bacterium glucosidase, Uniparc reference UPI0003DFF0DA, Uniprot reference W0FLD1.

[0893] SEQ ID NO: 745: Codon optimised DNA encoding 6×His Bacteroides xylanisolvens glucosidase, Uniparc reference UPI0002D3D994, Uniprot reference W6P9J9.

[0894] SEQ ID NO: 746: Codon optimised DNA encoding 6×His Fusarium oxysporum glucosidase, Uniparc reference UPI00042F8016, Uniprot reference X0HA35.

[0895] SEQ ID NO: 747: Codon optimised DNA encoding 6×His Sorghum bicolor glucosidase, Uniparc reference UPI00000A7F40, Uniprot reference Q41290.

[0896] SEQ ID NO: 748: Codon optimised DNA encoding 6×His Clostridium cellulovorans glucosidase, Uniparc reference UPI000050B701, Uniprot reference Q53EH2.

[0897] SEQ ID NO: 749: Codon optimised DNA encoding 6×His Dacryopinax primogenitus glucosidase, Uniparc reference UPI0002C29EAA, Uniprot reference M5G1U5.

[0898] SEQ ID NO: 750: Codon optimised DNA encoding 6×His Rhizobium radiobacter glucosidase, Uniparc reference UPI0000126913, Uniprot reference P27034.

[0899] SEQ ID NO: 751: Codon optimised DNA encoding 6×His Phanerochaete chrysosporium glucosidase, Uniparc reference UPI00006E0CC7, Uniprot reference Q25BW5A.

[0900] SEQ ID NO: 752: Codon optimised DNA encoding 6×His Enterobacter agglomerans glucosidase, Uniparc reference UPI0000126905, Uniprot reference Q59437.

[0901] SEQ ID NO: 753: Codon optimised DNA encoding 6×His Candidatus microthrix glucosidase, Uniparc reference UPI00033056C4, Uniprot reference R4YWU4.

[0902] SEQ ID NO: 754: Codon optimised DNA encoding 6×His Chthonomonas calidirosea glucosidase, Uniparc reference UPI0003427F29, Uniprot reference S0ET37.

[0903] SEQ ID NO: 755: Codon optimised DNA encoding 6×His Acholeplasma brassicae glucosidase, Uniparc reference UPI0003B04208, Uniprot reference U4KMR7.

[0904] SEQ ID NO: 756: Codon optimised DNA encoding 6×His Chania multitudinisentens glucosidase, Uniparc reference UPI0003E13BD3, Uniprot reference W0LJ28.

[0905] SEQ ID NO: 757: Codon optimised DNA encoding 6×His Gibberella moniliformis glucosidase, Uniparc reference UPI0003ECCF80, Uniprot reference W7MW98.

[0906] SEQ ID NO: 758: Codon optimised DNA encoding 6×His Fusarium oxysporum glucosidase, Uniparc reference UPI0004305856, Uniprot reference X0I2U8.

[0907] SEQ ID NO: 759: Codon optimised DNA encoding 6×His Phanerochaete chrysosporium glucosidase, Uniparc reference UPI00006E0CC7, Uniprot reference Q25BW5B.

[0908] SEQ ID NO: 760: Codon optimised DNA encoding 6×His Secale cereale glucosidase, Uniparc reference UPI00000A7EFC, Uniprot reference Q9FYS3.

[0909] SEQ ID NO: 761: Codon optimised DNA encoding 6×His Anoxybacillus gonensis glucosidase, Uniparc reference UPI0002C0273E, Uniprot reference M5QUM2.

[0910] SEQ ID NO: 762: Codon optimised DNA encoding 6×His Thermotoga maritima glucosidase, Uniparc reference UPI0000126906, Uniprot reference Q08638A.

[0911] SEQ ID NO: 763: Codon optimised DNA encoding 6×His Rhodospirillum rubrum glucosidase, Uniparc reference UPI00003C2ACC, Uniprot reference Q2RP51.

[0912] SEQ ID NO: 764: Codon optimised DNA encoding 6×His Thermotoga neapolitana glucosidase, Uniparc reference UPI00000B9013, Uniprot reference Q60038.

[0913] SEQ ID NO: 765: Codon optimised DNA encoding 6×His Candidatus microthrix glucosidase, Uniparc reference UPI00032F466E, Uniprot reference R4Z6M6.

[0914] SEQ ID NO: 766: Codon optimised DNA encoding 6×His Ruminiclostridium cellobioparum glucosidase, Uniparc reference UPI00032874EC, Uniprot reference S0FPI8.

[0915] SEQ ID NO: 767: Codon optimised DNA encoding 6×His Acholeplasma brassicae glucosidase, Uniparc reference UPI0003B0490C, Uniprot reference U4KMV4.

[0916] SEQ ID NO: 768: Codon optimised DNA encoding 6×His Klebsiella pneumoniae glucosidase, Uniparc reference UPI0003DB6EFD, Uniprot reference W1BBP5.

[0917] SEQ ID NO: 769: Codon optimised DNA encoding 6×His Enterobacter sp. glucosidase, Uniparc reference UPI0003ED0A8E, Uniprot reference W7P6Y1.

[0918] SEQ ID NO: 770: Codon optimised DNA encoding 6×His Rhizoctonia solani glucosidase, Uniparc reference UPI00045BB507, Uniprot reference X8JI88.

[0919] SEQ ID NO: 771: Codon optimised DNA encoding 6×His Homo sapiens glucosidase, Uniparc reference UPI0000072C73, Uniprot reference Q9H227.

[0920] SEQ ID NO: 772: Codon optimised DNA encoding 6×His uncultured bacterium glucosidase, Uniparc reference UPI0000DD5304, Uniprot reference Q0GMU3.

[0921] SEQ ID NO: 773: Codon optimised DNA encoding 6×His Thermoanaerobacter thermohydrosulfuricus glucosidase, Uniparc reference UPI0002CA9E4F, Uniprot reference M8CQD9.

[0922] SEQ ID NO: 774: Codon optimised DNA encoding 6×His Stigmatella aurantiaca glucosidase, Uniparc reference UPI0000E297D8, Uniprot reference Q091M8.

[0923] SEQ ID NO: 775: Codon optimised DNA encoding 6×His Burkholderia thailandensis glucosidase, Uniparc reference UPI00006676B1, Uniprot reference Q2T7C9.

[0924] SEQ ID NO: 776: Codon optimised DNA encoding 6×His Yersinia pseudotuberculosis glucosidase, Uniparc reference UPI0000426E8F, Uniprot reference Q665S2.

[0925] SEQ ID NO: 777: Codon optimised DNA encoding 6×His Togninia minima glucosidase, Uniparc reference UPI00032BF71F, Uniprot reference R8BQM8.

[0926] SEQ ID NO: 778: Codon optimised DNA encoding 6×His Ophiostoma piceae glucosidase, Uniparc reference UPI00035213C2, Uniprot reference S3BVJ8.

[0927] SEQ ID NO: 779: Codon optimised DNA encoding 6×His Pyronema omphalodes glucosidase, Uniparc reference UPI0003B117B7, Uniprot reference U4LIL3.

[0928] SEQ ID NO: 780: Codon optimised DNA encoding 6×His Ogataea parapolymorpha glucosidase, Uniparc reference UPI0001F76D1F, Uniprot reference W1QIW0.

[0929] SEQ ID NO: 781: Codon optimised DNA encoding 6×His Hymenobacter swuensis glucosidase, Uniparc reference UPI0003F2059F, Uniprot reference W8F6U0.

[0930] SEQ ID NO: 782: Codon optimised DNA encoding 6×His Flavobacterium johnsoniae glucosidase, Uniparc reference UPI00006E8E1B, Uniprot reference A5FEF5.

[0931] SEQ ID NO: 783: Codon optimised DNA encoding 6×His Paenibacillus polymyxa glucosidase, Uniparc reference UPI000012690B, Uniprot reference P22505.

[0932] SEQ ID NO: 784: Codon optimised DNA encoding 6×His Oryza sativa glucosidase, Uniparc reference UPI000009D014, Uniprot reference Q8L7J2.

[0933] SEQ ID NO: 785: Codon optimised DNA encoding 6×His Oryza sativa glucosidase, Uniparc reference UPI0000E580F2, Uniprot reference B8AVF0.

[0934] SEQ ID NO: 786: Codon optimised DNA encoding 6×His Nannochloris glucosidase, Uniparc reference UPI000EA7F16F, Uniprot reference A0A452CSM4.

[0935] SEQ ID NO: 787: Codon optimised DNA encoding 6×His Halothermothrix orenii glucosidase, Uniparc reference UPI00006AE508, Uniprot reference B8CYA8.

[0936] SEQ ID NO: 788: Codon optimised DNA encoding 6×His Neurospora crassa glucosidase, Uniparc reference UPI000018B2B4, Uniprot reference Q7RWP2.

[0937] SEQ ID NO: 789: Codon optimised DNA encoding 6×His Micrococcus antarcticus glucosidase, Uniparc reference UPI000192BB5F, Uniprot reference B9V8P5.

[0938] SEQ ID NO: 790: Codon optimised DNA encoding 6×His Exiguobacterium antarcticum glucosidase, Uniparc reference UPI000285E79E, Uniprot reference K0A8J9.

[0939] SEQ ID NO: 791: Codon optimised DNA encoding 6×His Thermus thermophilus glucosidase, Uniparc reference UPI00000BEB61, Uniprot reference Q9RA61.

[0940] SEQ ID NO: 792: Codon optimised DNA encoding 6×His Trichoderma harzianum glucosidase, Uniparc reference UPI00078BF747, Uniprot reference A0A2T4AR08.

[0941] SEQ ID NO: 793: Codon optimised DNA encoding 6×His Hypocrea jecorina glucosidase, Uniparc reference UPI000006AA61, Uniprot reference Q12715.

[0942] SEQ ID NO: 794: Codon optimised DNA encoding 6×His Streptomyces sp. glucosidase, Uniparc reference UPI00000B411B, Uniprot reference Q59976.

[0943] SEQ ID NO: 795: Codon optimised DNA encoding 6×His Streptococcus pyogenes glucosidase, Uniparc reference UPI00000C7E56, Uniprot reference Q99YP9.

[0944] SEQ ID NO: 796: Codon optimised DNA encoding 6×His Trifolium repens glucosidase, Uniparc reference UPI000012691B, Uniprot reference P26205.

[0945] SEQ ID NO: 797: Codon optimised DNA encoding 6×His Talaromyces emersonii glucosidase, Uniparc reference UPI000006C8FF, Uniprot reference Q8TGI8.

[0946] SEQ ID NO: 798: Codon optimised DNA encoding 6×His Hungateiclostridium thermocellum glucosidase, Uniparc reference UPI0000126903, Uniprot reference P26208.

[0947] SEQ ID NO: 799: Codon optimised DNA encoding 6×His Lactobacillus plantarum glucosidase, Uniparc reference UPI000219FE3E, Uniprot reference F9ULH8.

[0948] SEQ ID NO: 800: Codon optimised DNA encoding 6×His Agrobacterium tumefaciens glucosidase, Uniparc reference UPI0003F2033A, Uniprot reference A0A2I4PGZ0.

[0949] SEQ ID NO: 801: Amino acid sequence for Bifidobacterium actinocoloniiforme glucosidase, Uniparc reference UPI000503B26C, Uniprot reference A0A086YYS8.

[0950] SEQ ID NO: 802: Amino acid sequence for Bifidobacterium psychraerophilum glucosidase, Uniparc reference UPI000502B461, Uniprot reference A0A087CD28.

[0951] SEQ ID NO: 803: Amino acid sequence for Penicillium italicum glucosidase, Uniparc reference UPI00052B8681, Uniprot reference A0A0A2K704.

[0952] SEQ ID NO: 804: Amino acid sequence for Microbacterium trichothecenolyticum glucosidase, Uniparc reference UPI0005ED0AE6, Uniprot reference A0A0M2HDB3.

[0953] SEQ ID NO: 805: Amino acid sequence for uncultured Clostridium sp. glucosidase, Uniparc reference UPI0008206F38, Uniprot reference A0A1C6I6A3.

[0954] SEQ ID NO: 806: Amino acid sequence for Bifiguratus adelaidae glucosidase, Uniparc reference UPI000BC57319, Uniprot reference A0A261Y7Q8.

[0955] SEQ ID NO: 807: Amino acid sequence for Paenibacillus thiaminolyticus glucosidase, Uniparc reference UPI000B3B73CD, Uniprot reference A0A378ZIK3.

[0956] SEQ ID NO: 808: Amino acid sequence for Microbacterium lemovicicum glucosidase, Uniparc reference UPI000F8FB9BA, Uniprot reference A0A3S9WE68.

[0957] SEQ ID NO: 809: Amino acid sequence for Rhodococcus erythropolis glucosidase, Uniparc reference UPI00019923DC, Uniprot reference C1A1N4.

[0958] SEQ ID NO: 810: Amino acid sequence for Cutibacterium avidum glucosidase, Uniparc reference UPI00022C19B5, Uniprot reference G4CZU8.

[0959] SEQ ID NO: 811: Amino acid sequence for Clavibacter michiganensis glucosidase, Uniparc reference UPI0002C5A938, Uniprot reference M5BD10.

[0960] SEQ ID NO: 812: Amino acid sequence for Microbacterium sp. glucosidase, Uniparc reference UPI0003DE3509, Uniprot reference W0ZC23.

[0961] SEQ ID NO: 813: Amino acid sequence for Bifidobacterium actinocoloniiforme glucosidase, Uniparc reference UPI000507F38A, Uniprot reference A0A086YZL4.

[0962] SEQ ID NO: 814: Amino acid sequence for Bifidobacterium reuteri glucosidase, Uniparc reference UPI000506119A, Uniprot reference A0A087CR26.

[0963] SEQ ID NO: 815: Amino acid sequence for Propionibacterium freudenreichii glucosidase, Uniparc reference UPI0005A5CAC2, Uniprot reference A0A0A8RX48.

[0964] SEQ ID NO: 816: Amino acid sequence for Microbacterium hydrocarbonoxydans glucosidase, Uniparc reference UPI0005EC18F9, Uniprot reference A0A0M2HRU0.

[0965] SEQ ID NO: 817: Amino acid sequence for Pseudonocardia sp. glucosidase, Uniparc reference UPI00094B6D3B, Uniprot reference A0A1Q8KSV8.

[0966] SEQ ID NO: 818: Amino acid sequence for Tuber aestivum glucosidase, Uniparc reference UPI000BC13DF0, Uniprot reference A0A292PKV5.

[0967] SEQ ID NO: 819: Amino acid sequence for Propionibacterium australiense glucosidase, Uniparc reference UPI000E5B4EA5, Uniprot reference A0A383S7A9.

[0968] SEQ ID NO: 820: Amino acid sequence for Fusarium sp. glucosidase, Uniparc reference UPI001004B2C9, Uniprot reference A0A428T6E2.

[0969] SEQ ID NO: 821: Amino acid sequence for Nectria haematococca glucosidase, Uniparc reference UPI0001B69B5C, Uniprot reference C7ZBV0.

[0970] SEQ ID NO: 822: Amino acid sequence for Actinoplanes sp. glucosidase, Uniparc reference UPI00023EBB15, Uniprot reference G8S3E7.

[0971] SEQ ID NO: 823: Amino acid sequence for Streptomyces fulvissimus glucosidase, Uniparc reference UPI0003289BC6, Uniprot reference N0CMW2.

[0972] SEQ ID NO: 824: Amino acid sequence for Pestalotiopsis fici glucosidase, Uniparc reference UPI0003E02BF1, Uniprot reference W3WXF1.

[0973] SEQ ID NO: 825: Amino acid sequence for Bifidobacterium bohemicum glucosidase, Uniparc reference UPI0005025F2E, Uniprot reference A0A086ZGP0.

[0974] SEQ ID NO: 826: Amino acid sequence for Bifidobacterium saeculare glucosidase, Uniparc reference UPI0005084E52, Uniprot reference A0A087D0Q2.

[0975] SEQ ID NO: 827: Amino acid sequence for Bionectria ochroleuca glucosidase, Uniparc reference UPI00059674D6, Uniprot reference A0A0B7JW24.

[0976] SEQ ID NO: 828: Amino acid sequence for Bifidobacterium pseudocatenulatum glucosidase, Uniparc reference UPI0006C6D6F9, Uniprot reference A0A174AU04.

[0977] SEQ ID NO: 829: Amino acid sequence for Pseudonocardia sp. glucosidase, Uniparc reference UPI00094ABAB3, Uniprot reference A0A1Q8LPB4.

[0978] SEQ ID NO: 830: Amino acid sequence for bacterium glucosidase, Uniparc reference UPI000CB49A0B, Uniprot reference A0A2H5Z8Q4.

[0979] SEQ ID NO: 831: Amino acid sequence for Coleophoma crateriformis glucosidase, Uniparc reference UPI000E38A995, Uniprot reference A0A3D8Q771.

[0980] SEQ ID NO: 832: Amino acid sequence for Arthrobotrys oligospora glucosidase, Uniparc reference UPI001102DFA3, Uniprot reference A0A4Z0Y5Y8.

[0981] SEQ ID NO: 833: Amino acid sequence for Pyrenophora teres glucosidase, Uniparc reference UPI0001ECDCBD, Uniprot reference E3RFS2.

[0982] SEQ ID NO: 834: Amino acid sequence for Gordonia polyisoprenivorans glucosidase, Uniparc reference UPI00024F2A26, Uniprot reference H6MTQ7.

[0983] SEQ ID NO: 835: Amino acid sequence for Stigmatella aurantiaca glucosidase, Uniparc reference UPI0000E28E5D, Uniprot reference Q08S21.

[0984] SEQ ID NO: 836: Amino acid sequence for Pestalotiopsis fici glucosidase, Uniparc reference UPI0003E03A52, Uniprot reference W3WZ03.

[0985] SEQ ID NO: 837: Amino acid sequence for Bifidobacterium magnum glucosidase, Uniparc reference UPI0003B7B6EE, Uniprot reference A0A087BEN9.

[0986] SEQ ID NO: 838: Amino acid sequence for Bifidobacterium stellenboschense glucosidase, Uniparc reference UPI000503F283, Uniprot reference A0A087DFL8.

[0987] SEQ ID NO: 839: Amino acid sequence for Bionectria ochroleuca glucosidase, Uniparc reference UPI00059673EE, Uniprot reference A0A0B7K316.

[0988] SEQ ID NO: 840: Amino acid sequence for Hungatella hathewayi glucosidase, Uniparc reference UPI0006C069F3, Uniprot reference A0A174LVE3.

[0989] SEQ ID NO: 841: Amino acid sequence for Mycetocola reblochoni glucosidase, Uniparc reference UPI00097EB800, Uniprot reference A0A1R4J2F9.

[0990] SEQ ID NO: 842: Amino acid sequence for Nonomuraea sp. glucosidase, Uniparc reference UPI0009ABD7B3, Uniprot reference A0A2P91X34.

[0991] SEQ ID NO: 843: Amino acid sequence for Coleophoma crateriformis glucosidase, Uniparc reference UPI000E391DF6, Uniprot reference A0A3D8T9C2.

[0992] SEQ ID NO: 844: Amino acid sequence for Paenarthrobacter aurescens glucosidase, Uniparc reference UPI0000EC83AE, Uniprot reference A1R2K1.

[0993] SEQ ID NO: 845: Amino acid sequence for Kitasatospora setae glucosidase, Uniparc reference UPI0001F21F08, Uniprot reference E4N4F6.

[0994] SEQ ID NO: 846: Amino acid sequence for Nocardiopsis alba glucosidase, Uniparc reference UPI00027E251E, Uniprot reference J7L3Z8.

[0995] SEQ ID NO: 847: Amino acid sequence for Rhodococcus jostii glucosidase, Uniparc reference UPI0000DBA338, Uniprot reference Q0SCI4.

[0996] SEQ ID NO: 848: Amino acid sequence for uncultured microorganism glucosidase, Uniparc reference UPI0003EC942D, Uniprot reference W5X324.

[0997] SEQ ID NO: 849: Amino acid sequence for Bifidobacterium merycicum glucosidase, Uniparc reference UPI0005084B63, Uniprot reference A0A087BJ88.

[0998] SEQ ID NO: 850: Amino acid sequence for Bifidobacterium scardovii glucosidase, Uniparc reference UPI0005018B5B, Uniprot reference A0A087DGT3.

[0999] SEQ ID NO: 851: Amino acid sequence for Fusarium oxysporum glucosidase, Uniparc reference UPI00021EC81E, Uniprot reference A0A0C4DJL4.

[1000] SEQ ID NO: 852: Amino acid sequence for Paraphaeosphaeria sporulosa glucosidase, Uniparc reference UPI0007CE199C, Uniprot reference A0A177BXH1.

[1001] SEQ ID NO: 853: Amino acid sequence for Clostridium oryzae glucosidase, Uniparc reference UPI0009A50F8D, Uniprot reference A0A1V4IY77.

[1002] SEQ ID NO: 854: Amino acid sequence for Corynespora cassiicola glucosidase, Uniparc reference UPI000D22F5EB, Uniprot reference A0A2T2N4T6.

[1003] SEQ ID NO: 855: Amino acid sequence for Choiromyces venosus glucosidase, Uniparc reference UPI000F7332F8, Uniprot reference A0A3N4J9R6.

[1004] SEQ ID NO: 856: Amino acid sequence for Saccharopolyspora erythraea glucosidase, Uniparc reference UPI0000F550EC, Uniprot reference A4F982.

[1005] SEQ ID NO: 857: Amino acid sequence for Streptomyces venezuelae glucosidase, Uniparc reference UPI000204906E, Uniprot reference F2R0Y2.

[1006] SEQ ID NO: 858: Amino acid sequence for Arthrobacter sp. glucosidase, Uniparc reference UPI00027DFDB0, Uniprot reference J7LVB1.

[1007] SEQ ID NO: 859: Amino acid sequence for Thermobrachium celere glucosidase, Uniparc reference UPI00033420CB, Uniprot reference R7RMQ6.

[1008] SEQ ID NO: 860: Amino acid sequence for Drechslerella stenobrocha glucosidase, Uniparc reference UPI0003EA65B9, Uniprot reference W7HT93.

[1009] SEQ ID NO: 861: Amino acid sequence for Bifidobacterium minimum glucosidase, Uniparc reference UPI0003B38542, Uniprot reference A0A087BMR5.

[1010] SEQ ID NO: 862: Amino acid sequence for Bifidobacterium thermacidophilum glucosidase, Uniparc reference UPI000409825E, Uniprot reference A0A087E3K8.

[1011] SEQ ID NO: 863: Amino acid sequence for Verruconis gallopava glucosidase, Uniparc reference UPI0005C0229F, Uniprot reference A0A0D1X9G0.

[1012] SEQ ID NO: 864: Amino acid sequence for Stagonospora sp. glucosidase, Uniparc reference UPI0007CECAD5, Uniprot reference A0A178AER7.

[1013] SEQ ID NO: 865: Amino acid sequence for Firmicutes bacterium glucosidase, Uniparc reference UPI0009D58C49, Uniprot reference A0A1V6ALF9.

[1014] SEQ ID NO: 866: Amino acid sequence for Tuber borchii glucosidase, Uniparc reference UPI000D50EED1, Uniprot reference A0A2T6ZNU7.

[1015] SEQ ID NO: 867: Amino acid sequence for Morchella conica glucosidase, Uniparc reference UPI000F728167, Uniprot reference A0A3N4KUR3.

[1016] SEQ ID NO: 868: Amino acid sequence for Dictyoglomus thermophilum glucosidase, Uniparc reference UPI00018152EB, Uniprot reference B5YC96.

[1017] SEQ ID NO: 869: Amino acid sequence for Microlunatus phosphovorus glucosidase, Uniparc reference UPI000210C01C, Uniprot reference F5XL24.

[1018] SEQ ID NO: 870: Amino acid sequence for Bifidobacterium asteroides glucosidase, Uniparc reference UPI00028BB1B6, Uniprot reference K4IPD2.

[1019] SEQ ID NO: 871: Amino acid sequence for Dactylellina haptotyla glucosidase, Uniparc reference UPI00035AE576, Uniprot reference S8BQ60.

[1020] SEQ ID NO: 872: Amino acid sequence for Fusarium oxysporum glucosidase, Uniparc reference UPI0003F2D7DD, Uniprot reference W9JF54.

[1021] SEQ ID NO: 873: Amino acid sequence for Bifidobacterium longum glucosidase, Uniparc reference UPI0005067FF9, Uniprot reference A0A087BRY2.

[1022] SEQ ID NO: 874: Amino acid sequence for Bifidobacterium tsurumiense glucosidase, Uniparc reference UPI0004105550, Uniprot reference A0A087EK01.

[1023] SEQ ID NO: 875: Amino acid sequence for Exophiala spinifera glucosidase, Uniparc reference UPI0005BF9DF9, Uniprot reference A0A0D1YG50.

[1024] SEQ ID NO: 876: Amino acid sequence for Pyrenochaeta sp. glucosidase, Uniparc reference UPI0007CE7DBA, Uniprot reference A0A178E511.

[1025] SEQ ID NO: 877: Amino acid sequence for Firmicutes bacterium glucosidase, Uniparc reference UPI0009CDA3F1, Uniprot reference A0A1V6FLK2.

[1026] SEQ ID NO: 878: Amino acid sequence for Cadophora sp. glucosidase, Uniparc reference UPI000D5A9E68, Uniprot reference A0A2V1B599.

[1027] SEQ ID NO: 879: Amino acid sequence for Morchella conica glucosidase, Uniparc reference UPI000F7330CD, Uniprot reference A0A3N4L4M3.

[1028] SEQ ID NO: 880: Amino acid sequence for Bifidobacterium animalis glucosidase, Uniparc reference UPI000189C68F, Uniprot reference B8DV42.

[1029] SEQ ID NO: 881: Amino acid sequence for Treponema azotonutricium glucosidase, Uniparc reference UPI00020ED2C9, Uniprot reference F5YGD5.

[1030] SEQ ID NO: 882: Amino acid sequence for Acidipropionibacterium acidipropionici glucosidase, Uniparc reference UPI0002988429, Uniprot reference K7S0E5.

[1031] SEQ ID NO: 883: Amino acid sequence for Salinispira pacifica glucosidase, Uniparc reference UPI0003D93613, Uniprot reference V5WNB6.

[1032] SEQ ID NO: 884: Amino acid sequence for Bifidobacterium mongoliense glucosidase, Uniparc reference UPI000503588E, Uniprot reference A0A087C760.

[1033] SEQ ID NO: 885: Amino acid sequence for Bifidobacterium indicum glucosidase, Uniparc reference UPI000499F5D1, Uniprot reference A0A087VUW4.

[1034] SEQ ID NO: 886: Amino acid sequence for Brachyspira suanatina glucosidase, Uniparc reference UPI000659257F, Uniprot reference A0A0G4K5C2.

[1035] SEQ ID NO: 887: Amino acid sequence for Phialocephala scopiformis glucosidase, Uniparc reference UPI0007F2FD44, Uniprot reference A0A194X8Q9.

[1036] SEQ ID NO: 888: Amino acid sequence for uncultured Microbacterium sp. glucosidase, Uniparc reference UPI000A2BB4A5, Uniprot reference A0A1Y5P895.

[1037] SEQ ID NO: 889: Amino acid sequence for Periconia macrospinosa glucosidase, Uniparc reference UPI000D5BC9BD, Uniprot reference A0A2V1E673.

[1038] SEQ ID NO: 890: Amino acid sequence for Arthrobacter ulcerisalmonis glucosidase, Uniparc reference UPI000F3DC30B, Uniprot reference A0A3P5WTE2.

[1039] SEQ ID NO: 891: Amino acid sequence for Pseudarthrobacter chlorophenolicus glucosidase, Uniparc reference UPI0001664880, Uniprot reference B8H9A2.

[1040] SEQ ID NO: 892: Amino acid sequence for Haloplasma contractile glucosidase, Uniparc reference UPI0002120C63, Uniprot reference F7PUF3.

[1041] SEQ ID NO: 893: Amino acid sequence for Cochliobolus heterostrophus glucosidase, Uniparc reference UPI0002B73341, Uniprot reference M2ULB2.

[1042] SEQ ID NO: 894: Amino acid sequence for Microbacterium sp. glucosidase, Uniparc reference UPI0003DE58EA, Uniprot reference W0Z818.

[1043] SEQ ID NO: 895: Codon optimised DNA encoding 6×His Bifidobacterium actinocoloniiforme glucosidase, Uniparc reference UPI000503B26C, Uniprot reference A0A086YYS8.

[1044] SEQ ID NO: 896: Codon optimised DNA encoding 6×His Bifidobacterium psychraerophilum glucosidase, Uniparc reference UPI000502B461, Uniprot reference A0A087CD28.

[1045] SEQ ID NO: 897: Codon optimised DNA encoding 6×His Penicillium italicum glucosidase, Uniparc reference UPI00052B8681, Uniprot reference A0A0A2K704.

[1046] SEQ ID NO: 898: Codon optimised DNA encoding 6×His Microbacterium trichothecenolyticum glucosidase, Uniparc reference UPI0005ED0AE6, Uniprot reference A0A0M2HDB3.

[1047] SEQ ID NO: 899: Codon optimised DNA encoding 6×His uncultured Clostridium sp. glucosidase, Uniparc reference UPI0008206F38, Uniprot reference A0A1C616A3.

[1048] SEQ ID NO: 900: Codon optimised DNA encoding 6×His Bifiguratus adelaidae glucosidase, Uniparc reference UPI000BC57319, Uniprot reference A0A261Y7Q8.

[1049] SEQ ID NO: 901: Codon optimised DNA encoding 6×His Paenibacillus thiaminolyticus glucosidase, Uniparc reference UPI000B3B73CD, Uniprot reference A0A378ZIK3.

[1050] SEQ ID NO: 902: Codon optimised DNA encoding 6×His Microbacterium lemovicicum glucosidase, Uniparc reference UPI000F8FB9BA, Uniprot reference A0A3S9WE68.

[1051] SEQ ID NO: 903: Codon optimised DNA encoding 6×His Rhodococcus erythropolis glucosidase, Uniparc reference UPI00019923DC, Uniprot reference C1A1N4.

[1052] SEQ ID NO: 904: Codon optimised DNA encoding 6×His Cutibacterium avidum glucosidase, Uniparc reference UPI00022C19B5, Uniprot reference G4CZU8.

[1053] SEQ ID NO: 905: Codon optimised DNA encoding 6×His Clavibacter michiganensis glucosidase, Uniparc reference UPI0002C5A938, Uniprot reference M5BD10.

[1054] SEQ ID NO: 906: Codon optimised DNA encoding 6×His Microbacterium sp. glucosidase, Uniparc reference UPI0003DE3509, Uniprot reference W0ZC23.

[1055] SEQ ID NO: 907: Codon optimised DNA encoding 6×His Bifidobacterium actinocoloniiforme glucosidase, Uniparc reference UPI000507F38A, Uniprot reference A0A086YZL4.

[1056] SEQ ID NO: 908: Codon optimised DNA encoding 6×His Bifidobacterium reuteri glucosidase, Uniparc reference UPI000506119A, Uniprot reference A0A087CR26.

[1057] SEQ ID NO: 909: Codon optimised DNA encoding 6×His Propionibacterium freudenreichii glucosidase, Uniparc reference UPI0005A5CAC2, Uniprot reference A0A0A8RX48.

[1058] SEQ ID NO: 910: Codon optimised DNA encoding 6×His Microbacterium hydrocarbonoxydans glucosidase, Uniparc reference UPI0005EC18F9, Uniprot reference A0A0M2HRU0.

[1059] SEQ ID NO: 911: Codon optimised DNA encoding 6×His Pseudonocardia sp. glucosidase, Uniparc reference UPI00094B6D3B, Uniprot reference A0A1Q8KSV8.

[1060] SEQ ID NO: 912: Codon optimised DNA encoding 6×His Tuber aestivum glucosidase, Uniparc reference UPI000BC13DF0, Uniprot reference A0A292PKV5.

[1061] SEQ ID NO: 913: Codon optimised DNA encoding 6×His Propionibacterium australiense glucosidase, Uniparc reference UPI000E5B4EA5, Uniprot reference A0A383S7A9.

[1062] SEQ ID NO: 914: Codon optimised DNA encoding 6×His Fusarium sp. glucosidase, Uniparc reference UPI001004B2C9, Uniprot reference A0A428T6E2.

[1063] SEQ ID NO: 915: Codon optimised DNA encoding 6×His Nectria haematococca glucosidase, Uniparc reference UPI0001B69B5C, Uniprot reference C7ZBV0.

[1064] SEQ ID NO: 916: Codon optimised DNA encoding 6×His Actinoplanes sp. glucosidase, Uniparc reference UPI00023EBB15, Uniprot reference G8S3E7.

[1065] SEQ ID NO: 917: Codon optimised DNA encoding 6×His Streptomyces fulvissimus glucosidase, Uniparc reference UPI0003289BC6, Uniprot reference N0CMW2.

[1066] SEQ ID NO: 918: Codon optimised DNA encoding 6×His Pestalotiopsis fici glucosidase, Uniparc reference UPI0003E02BF1, Uniprot reference W3WXF1.

[1067] SEQ ID NO: 919: Codon optimised DNA encoding 6×His Bifidobacterium bohemicum glucosidase, Uniparc reference UPI0005025F2E, Uniprot reference A0A086ZGP0.

[1068] SEQ ID NO: 920: Codon optimised DNA encoding 6×His Bifidobacterium saeculare glucosidase, Uniparc reference UPI0005084E52, Uniprot reference A0A087D0Q2.

[1069] SEQ ID NO: 921: Codon optimised DNA encoding 6×His Bionectria ochroleuca glucosidase, Uniparc reference UPI00059674D6, Uniprot reference A0A0B7JW24.

[1070] SEQ ID NO: 922: Codon optimised DNA encoding 6×His Bifidobacterium pseudocatenulatum glucosidase, Uniparc reference UPI0006C6D6F9, Uniprot reference A0A174AU04.

[1071] SEQ ID NO: 923: Codon optimised DNA encoding 6×His Pseudonocardia sp. glucosidase, Uniparc reference UPI00094ABAB3, Uniprot reference A0A1Q8LPB4.

[1072] SEQ ID NO: 924: Codon optimised DNA encoding 6×His bacterium glucosidase, Uniparc reference UPI000CB49A0B, Uniprot reference A0A2H5Z8Q4.

[1073] SEQ ID NO: 925: Codon optimised DNA encoding 6×His Coleophoma crateriformis glucosidase, Uniparc reference UPI000E38A995, Uniprot reference A0A3D8Q771.

[1074] SEQ ID NO: 926: Codon optimised DNA encoding 6×His Arthrobotrys oligospora glucosidase, Uniparc reference UPI001102DFA3, Uniprot reference A0A4Z0Y5Y8.

[1075] SEQ ID NO: 927: Codon optimised DNA encoding 6×His Pyrenophora teres glucosidase, Uniparc reference UPI0001ECDCBD, Uniprot reference E3RFS2.

[1076] SEQ ID NO: 928: Codon optimised DNA encoding 6×His Gordonia polyisoprenivorans glucosidase, Uniparc reference UPI00024F2A26, Uniprot reference H6MTQ7.

[1077] SEQ ID NO: 929: Codon optimised DNA encoding 6×His Stigmatella aurantiaca glucosidase, Uniparc reference UPI0000E28E5D, Uniprot reference Q08S21.

[1078] SEQ ID NO: 930: Codon optimised DNA encoding 6×His Pestalotiopsis fici glucosidase, Uniparc reference UPI0003E03A52, Uniprot reference W3WZ03.

[1079] SEQ ID NO: 931: Codon optimised DNA encoding 6×His Bifidobacterium magnum glucosidase, Uniparc reference UPI0003B7B6EE, Uniprot reference A0A087BEN9.

[1080] SEQ ID NO: 932: Codon optimised DNA encoding 6×His Bifidobacterium stellenboschense glucosidase, Uniparc reference UPI000503F283, Uniprot reference A0A087DFL8.

[1081] SEQ ID NO: 933: Codon optimised DNA encoding 6×His Bionectria ochroleuca glucosidase, Uniparc reference UPI00059673EE, Uniprot reference A0A0B7K316.

[1082] SEQ ID NO: 934: Codon optimised DNA encoding 6×His Hungatella hathewayi glucosidase, Uniparc reference UPI0006C069F3, Uniprot reference A0A174LVE3.

[1083] SEQ ID NO: 935: Codon optimised DNA encoding 6×His Mycetocola reblochoni glucosidase, Uniparc reference UPI00097EB800, Uniprot reference A0A1 R4J2F9.

[1084] SEQ ID NO: 936: Codon optimised DNA encoding 6×His Nonomuraea sp. glucosidase, Uniparc reference UPI0009ABD7B3, Uniprot reference A0A2P91X34.

[1085] SEQ ID NO: 937: Codon optimised DNA encoding 6×His Coleophoma crateriformis glucosidase, Uniparc reference UPI000E391DF6, Uniprot reference A0A3D8T9C2.

[1086] SEQ ID NO: 938: Codon optimised DNA encoding 6×His Paenarthrobacter aurescens glucosidase, Uniparc reference UPI0000EC83AE, Uniprot reference A1R2K1.

[1087] SEQ ID NO: 939: Codon optimised DNA encoding 6×His Kitasatospora setae glucosidase, Uniparc reference UPI0001F21F08, Uniprot reference E4N4F6.

[1088] SEQ ID NO: 940: Codon optimised DNA encoding 6×His Nocardiopsis alba glucosidase, Uniparc reference UPI00027E251E, Uniprot reference J7L3Z8.

[1089] SEQ ID NO: 941: Codon optimised DNA encoding 6×His Rhodococcus jostii glucosidase, Uniparc reference UPI0000DBA338, Uniprot reference Q0SCI4.

[1090] SEQ ID NO: 942: Codon optimised DNA encoding 6×His uncultured microorganism glucosidase, Uniparc reference UPI0003EC942D, Uniprot reference W5X324.

[1091] SEQ ID NO: 943: Codon optimised DNA encoding 6×His Bifidobacterium merycicum glucosidase, Uniparc reference UPI0005084B63, Uniprot reference A0A087BJ88.

[1092] SEQ ID NO: 944: Codon optimised DNA encoding 6×His Bifidobacterium scardovii glucosidase, Uniparc reference UPI0005018B5B, Uniprot reference A0A087DGT3.

[1093] SEQ ID NO: 945: Codon optimised DNA encoding 6×His Fusarium oxysporum glucosidase, Uniparc reference UPI00021EC81E, Uniprot reference A0A0C4DJL4.

[1094] SEQ ID NO: 946: Codon optimised DNA encoding 6×His Paraphaeosphaeria sporulosa glucosidase, Uniparc reference UPI0007CE199C, Uniprot reference A0A177BXH1.

[1095] SEQ ID NO: 947: Codon optimised DNA encoding 6×His Clostridium oryzae glucosidase, Uniparc reference UPI0009A50F8D, Uniprot reference A0A1V41Y77.

[1096] SEQ ID NO: 948: Codon optimised DNA encoding 6×His Corynespora cassiicola glucosidase, Uniparc reference UPI000D22F5EB, Uniprot reference A0A2T2N4T6.

[1097] SEQ ID NO: 949: Codon optimised DNA encoding 6×His Choiromyces venosus glucosidase, Uniparc reference UPI000F7332F8, Uniprot reference A0A3N4J9R6.

[1098] SEQ ID NO: 950: Codon optimised DNA encoding 6×His Saccharopolyspora erythraea glucosidase, Uniparc reference UPI0000F550EC, Uniprot reference A4F982.

[1099] SEQ ID NO: 951: Codon optimised DNA encoding 6×His Streptomyces venezuelae glucosidase, Uniparc reference UPI000204906E, Uniprot reference F2R0Y2.

[1100] SEQ ID NO: 952: Codon optimised DNA encoding 6×His Arthrobacter sp. glucosidase, Uniparc reference UPI00027DFDB0, Uniprot reference J7LVB1.

[1101] SEQ ID NO: 953: Codon optimised DNA encoding 6×His Thermobrachium celere glucosidase, Uniparc reference UPI00033420CB, Uniprot reference R7RMQ6.

[1102] SEQ ID NO: 954: Codon optimised DNA encoding 6×His Drechslerella stenobrocha glucosidase, Uniparc reference UPI0003EA65B9, Uniprot reference W7HT93.

[1103] SEQ ID NO: 955: Codon optimised DNA encoding 6×His Bifidobacterium minimum glucosidase, Uniparc reference UPI0003B38542, Uniprot reference A0A087BMR5.

[1104] SEQ ID NO: 956: Codon optimised DNA encoding 6×His Bifidobacterium thermacidophilum glucosidase, Uniparc reference UPI000409825E, Uniprot reference A0A087E3K8.

[1105] SEQ ID NO: 957: Codon optimised DNA encoding 6×His Verruconis gallopava glucosidase, Uniparc reference UPI0005C0229F, Uniprot reference A0A0D1X9G0.

[1106] SEQ ID NO: 958: Codon optimised DNA encoding 6×His Stagonospora sp. glucosidase, Uniparc reference UPI0007CECAD5, Uniprot reference A0A178AER7.

[1107] SEQ ID NO: 959: Codon optimised DNA encoding 6×His Firmicutes bacterium glucosidase, Uniparc reference UPI0009D58C49, Uniprot reference A0A1V6ALF9.

[1108] SEQ ID NO: 960: Codon optimised DNA encoding 6×His Tuber borchii glucosidase, Uniparc reference UPI000D50EED1, Uniprot reference A0A2T6ZNU7.

[1109] SEQ ID NO: 961: Codon optimised DNA encoding 6×His Morchella conica glucosidase, Uniparc reference UPI000F728167, Uniprot reference A0A3N4KUR3.

[1110] SEQ ID NO: 962: Codon optimised DNA encoding 6×His Dictyoglomus thermophilum glucosidase, Uniparc reference UPI00018152EB, Uniprot reference B5YC96.

[1111] SEQ ID NO: 963: Codon optimised DNA encoding 6×His Microlunatus phosphovorus glucosidase, Uniparc reference UPI000210C01C, Uniprot reference F5XL24.

[1112] SEQ ID NO: 964: Codon optimised DNA encoding 6×His Bifidobacterium asteroides glucosidase, Uniparc reference UPI00028BB1B6, Uniprot reference K4IPD2.

[1113] SEQ ID NO: 965: Codon optimised DNA encoding 6×His Dactylellina haptotyla glucosidase, Uniparc reference UPI00035AE576, Uniprot reference S8BQ60.

[1114] SEQ ID NO: 966: Codon optimised DNA encoding 6×His Fusarium oxysporum glucosidase, Uniparc reference UPI0003F2D7DD, Uniprot reference W9JF54.

[1115] SEQ ID NO: 967: Codon optimised DNA encoding 6×His Bifidobacterium longum glucosidase, Uniparc reference UPI0005067FF9, Uniprot reference A0A087BRY2.

[1116] SEQ ID NO: 968: Codon optimised DNA encoding 6×His Bifidobacterium tsurumiense glucosidase, Uniparc reference UPI0004105550, Uniprot reference A0A087EK01.

[1117] SEQ ID NO: 969: Codon optimised DNA encoding 6×His Exophiala spinifera glucosidase, Uniparc reference UPI0005BF9DF9, Uniprot reference A0A0D1YG50.

[1118] SEQ ID NO: 970: Codon optimised DNA encoding 6×His Pyrenochaeta sp. glucosidase, Uniparc reference UPI0007CE7DBA, Uniprot reference A0A178E511.

[1119] SEQ ID NO: 971: Codon optimised DNA encoding 6×His Firmicutes bacterium glucosidase, Uniparc reference UPI0009CDA3F1, Uniprot reference A0A1V6FLK2.

[1120] SEQ ID NO: 972: Codon optimised DNA encoding 6×His Cadophora sp. glucosidase, Uniparc reference UPI000D5A9E68, Uniprot reference A0A2V1B599.

[1121] SEQ ID NO: 973: Codon optimised DNA encoding 6×His Morchella conica glucosidase, Uniparc reference UPI000F7330CD, Uniprot reference A0A3N4L4M3.

[1122] SEQ ID NO: 974: Codon optimised DNA encoding 6×His Bifidobacterium animalis glucosidase, Uniparc reference UPI000189C68F, Uniprot reference B8DV42.

[1123] SEQ ID NO: 975: Codon optimised DNA encoding 6×His Treponema azotonutricium glucosidase, Uniparc reference UPI00020ED2C9, Uniprot reference F5YGD5.

[1124] SEQ ID NO: 976: Codon optimised DNA encoding 6×His Acidipropionibacterium acidipropionici glucosidase, Uniparc reference UPI0002988429, Uniprot reference K7S0E5.

[1125] SEQ ID NO: 977: Codon optimised DNA encoding 6×His Salinispira pacifica glucosidase, Uniparc reference UPI0003D93613, Uniprot reference V5WNB6.

[1126] SEQ ID NO: 978: Codon optimised DNA encoding 6×His Bifidobacterium mongoliense glucosidase, Uniparc reference UPI000503588E, Uniprot reference A0A087C760.

[1127] SEQ ID NO: 979: Codon optimised DNA encoding 6×His Bifidobacterium indicum glucosidase, Uniparc reference UPI000499F5D1, Uniprot reference A0A087VUW4.

[1128] SEQ ID NO: 980: Codon optimised DNA encoding 6×His Brachyspira suanatina glucosidase, Uniparc reference UPI000659257F, Uniprot reference A0A0G4K5C2.

[1129] SEQ ID NO: 981: Codon optimised DNA encoding 6×His Phialocephala scopiformis glucosidase, Uniparc reference UPI0007F2FD44, Uniprot reference A0A194X8Q9.

[1130] SEQ ID NO: 982: Codon optimised DNA encoding 6×His uncultured Microbacterium sp. glucosidase, Uniparc reference UPI000A2BB4A5, Uniprot reference A0A1Y5P895.

[1131] SEQ ID NO: 983: Codon optimised DNA encoding 6×His Periconia macrospinosa glucosidase, Uniparc reference UPI000D5BC9BD, Uniprot reference A0A2V1E673.

[1132] SEQ ID NO: 984: Codon optimised DNA encoding 6×His Arthrobacter ulcerisalmonis glucosidase, Uniparc reference UPI000F3DC30B, Uniprot reference A0A3P5WTE2.

[1133] SEQ ID NO: 985: Codon optimised DNA encoding 6×His Pseudarthrobacter chlorophenolicus glucosidase, Uniparc reference UPI0001664880, Uniprot reference B8H9A2.

[1134] SEQ ID NO: 986: Codon optimised DNA encoding 6×His Haloplasma contractile glucosidase, Uniparc reference UPI0002120C63, Uniprot reference F7PUF3.

[1135] SEQ ID NO: 987: Codon optimised DNA encoding 6×His Cochliobolus heterostrophus glucosidase, Uniparc reference UPI0002B73341, Uniprot reference M2ULB2.

[1136] SEQ ID NO: 988: Codon optimised DNA encoding 6×His Microbacterium sp. glucosidase, Uniparc reference UPI0003DE58EA, Uniprot reference W0Z818.

[1137] SEQ ID NO: 989: Amino acid sequence for Rhodothermus marinus rhamnosidase, Uniparc reference UPI0001A31108, Uniprot reference D0MFR0.

[1138] SEQ ID NO: 990: Amino acid sequence for Streptomyces bingchenggensis rhamnosidase, Uniparc reference UPI0001D90BFE, Uniprot reference D7C463.

[1139] SEQ ID NO: 991: Amino acid sequence for Spirosoma linguale rhamnosidase, Uniparc reference UPI0001A3AEAC, Uniprot reference D2QL60.

[1140] SEQ ID NO: 992: Amino acid sequence for Roseburia intestinalis rhamnosidase, Uniparc reference UPI0001CD6D48, Uniprot reference D4L2K8.

[1141] SEQ ID NO: 993: Amino acid sequence for Draconibacterium orientale rhamnosidase, Uniparc reference UPI000442EF22, Uniprot reference X5DG83.

[1142] SEQ ID NO: 994: Amino acid sequence for Catenulispora acidiphila rhamnosidase, Uniparc reference UPI00019E052B, Uniprot reference C7QC24.

[1143] SEQ ID NO: 995: Amino acid sequence for Bacteroides thetaiotaomicron rhamnosidase, Uniparc reference UPI000005ADE1, Uniprot reference Q8A916.

[1144] SEQ ID NO: 996: Amino acid sequence for Opitutus terrae rhamnosidase, Uniparc reference UPI000172B2E3, Uniprot reference B1ZRE4.

[1145] SEQ ID NO: 997: Amino acid sequence for Lachnoclostridium phytofermentans rhamnosidase, Uniparc reference UPI00015FE0BE, Uniprot reference A9KJP8.

[1146] SEQ ID NO: 998: Amino acid sequence for Rhodanobacter denitrificans rhamnosidase, Uniparc reference UPI00022DA3ED, Uniprot reference M4NH01.

[1147] SEQ ID NO: 999: Amino acid sequence for Prevotella ruminicola rhamnosidase, Uniparc reference UPI0001D07633, Uniprot reference D5ETD9.

[1148] SEQ ID NO: 1000: Amino acid sequence for Aspergillus terreus rhamnosidase, Uniparc reference UPI000259E388, Uniprot reference 10AZ41.

[1149] SEQ ID NO: 1001: Amino acid sequence for Brachybacterium faecium rhamnosidase, Uniparc reference UPI0001A461B6, Uniprot reference C7MA58.

[1150] SEQ ID NO: 1002: Amino acid sequence for Flavobacterium johnsoniae rhamnosidase, Uniparc reference UPI00006E4A0D, Uniprot reference A5FCH3.

[1151] SEQ ID NO: 1003: Amino acid sequence for Rahnella aquatilis rhamnosidase, Uniparc reference UPI000245C507, Uniprot reference H2IYR2.

[1152] SEQ ID NO: 1004: Amino acid sequence for Bifidobacterium moukalabense rhamnosidase, Uniparc reference UPI0003E4E711, Uniprot reference W4N6H0.

[1153] SEQ ID NO: 1005: Amino acid sequence for Enterococcus casseliflavus rhamnosidase, Uniparc reference UPI000353BD9F, Uniprot reference S4BBS9.

[1154] SEQ ID NO: 1006: Amino acid sequence for Geobacillus sp. rhamnosidase, Uniparc reference UPI0001789COD, Uniprot reference D3EED1.

[1155] SEQ ID NO: 1007: Amino acid sequence for Modestobacter marinus rhamnosidase, Uniparc reference UPI000260A2FE, Uniprot reference I4EYD9.

[1156] SEQ ID NO: 1008: Amino acid sequence for Pedobacter heparinus rhamnosidase, Uniparc reference UPI00019EF0EE, Uniprot reference C6XYM6.

[1157] SEQ ID NO: 1009: Amino acid sequence for Dyadobacter fermentans rhamnosidase, Uniparc reference UPI00019B5915, Uniprot reference C6VZL3.

[1158] SEQ ID NO: 1010: Amino acid sequence for Paenibacillus mucilaginosus rhamnosidase, Uniparc reference UPI000212C156, Uniprot reference F8FQQ3.

[1159] SEQ ID NO: 1011: Amino acid sequence for Paenibacillus sp. rhamnosidase, Uniparc reference UPI0003E2544D, Uniprot reference W4D866.

[1160] SEQ ID NO: 1012: Amino acid sequence for Bacteroides thetaiotaomicron rhamnosidase, Uniparc reference UPI000005ADD2, Uniprot reference A0A0P0FM19.

[1161] SEQ ID NO: 1013: Amino acid sequence for Chloroflexus aurantiacus rhamnosidase, Uniparc reference UPI00005BA60B, Uniprot reference A9WDK5.

[1162] SEQ ID NO: 1014: Amino acid sequence for Thermoclostridium stercorarium rhamnosidase, Uniparc reference UPI00000B098C, Uniprot reference Q9S3L0.

[1163] SEQ ID NO: 1015: Amino acid sequence for Bifidobacterium moukalabense rhamnosidase, Uniparc reference UPI0003E5C314, Uniprot reference W4N612.

[1164] SEQ ID NO: 1016: Amino acid sequence for Olsenella profusa rhamnosidase, Uniparc reference UPI0003AE032C, Uniprot reference U2USP4.

[1165] SEQ ID NO: 1017: Amino acid sequence for Kribbella flavida rhamnosidase, Uniparc reference UPI00019BDB13, Uniprot reference D2PMT5.

[1166] SEQ ID NO: 1018: Amino acid sequence for Caulobacter vibrioides rhamnosidase, Uniparc reference UPI00000C7226, Uniprot reference Q9A9K2.

[1167] SEQ ID NO: 1019: Amino acid sequence for Bacteroides thetaiotaomicron rhamnosidase, Uniparc reference UPI000005BA09, Uniprot reference Q8A076.

[1168] SEQ ID NO: 1020: Amino acid sequence for Rhodonellum psychrophilum rhamnosidase, Uniparc reference UPI0003745394, Uniprot reference U5BUY4.

[1169] SEQ ID NO: 1021: Amino acid sequence for Paenibacillus sp. rhamnosidase, Uniparc reference UPI0004F6D660, Uniprot reference A0A089M3T2.

[1170] SEQ ID NO: 1022: Amino acid sequence for Catenovulum agarivorans rhamnosidase, Uniparc reference UPI0003ED860D, Uniprot reference W7QMH5.

[1171] SEQ ID NO: 1023: Amino acid sequence for Zobellia galactanivorans rhamnosidase, Uniparc reference UPI000217D8B1, Uniprot reference G0L382.

[1172] SEQ ID NO: 1024: Amino acid sequence for Bacteroides thetaiotaomicron rhamnosidase, Uniparc reference UPI000005B845, Uniprot reference Q8A1H5.

[1173] SEQ ID NO: 1025: Amino acid sequence for Bacteroides xylanisolvens rhamnosidase, Uniparc reference UPI0001CD02E9, Uniprot reference D6CYE5.

[1174] SEQ ID NO: 1026: Amino acid sequence for Pseudarthrobacter chlorophenolicus rhamnosidase, Uniparc reference UPI00018E07C6, Uniprot reference B8HAH3.

[1175] SEQ ID NO: 1027: Amino acid sequence for Dictyoglomus thermophilum rhamnosidase, Uniparc reference UPI0001815896, Uniprot reference B5YC64.

[1176] SEQ ID NO: 1028: Amino acid sequence for Formosa agariphila rhamnosidase, Uniparc reference UPI00039231C1, Uniprot reference T2KPL4.

[1177] SEQ ID NO: 1029: Amino acid sequence for Rhodococcus jostii rhamnosidase, Uniparc reference UPI0000DBA6EB, Uniprot reference Q0S9T4.

[1178] SEQ ID NO: 1030: Amino acid sequence for Lactobacillus crispatus rhamnosidase, Uniparc reference UPI0001D10896, Uniprot reference D5GZ45.

[1179] SEQ ID NO: 1031: Amino acid sequence for Pedobacter heparinus rhamnosidase, Uniparc reference UPI0001B17DE6, Uniprot reference C6XVU2.

[1180] SEQ ID NO: 1032: Amino acid sequence for Spirosoma linguale rhamnosidase, Uniparc reference UPI0001A3C989, Uniprot reference D2QUA5.

[1181] SEQ ID NO: 1033: Amino acid sequence for Pedobacter heparinus rhamnosidase, Uniparc reference UPI00019EE3EB, Uniprot reference C6XU05.

[1182] SEQ ID NO: 1034: Amino acid sequence for Paenibacillus mucilaginosus rhamnosidase, Uniparc reference UPI0003432C11, Uniprot reference R9ULQ4.

[1183] SEQ ID NO: 1035: Amino acid sequence for Caulobacter segnis rhamnosidase, Uniparc reference UPI0001BC0C05, Uniprot reference D5VGD9.

[1184] SEQ ID NO: 1036: Amino acid sequence for Bacteroides cellulosilyticus rhamnosidase, Uniparc reference UPI0001969377, Uniprot reference E2N9B1.

[1185] SEQ ID NO: 1037: Amino acid sequence for Pedobacter heparinus rhamnosidase, Uniparc reference UPI0001B17C60, Uniprot reference C6Y153.

[1186] SEQ ID NO: 1038: Amino acid sequence for Formosa agariphila rhamnosidase, Uniparc reference UPI000571C0C2, Uniprot reference T2KNB2.

[1187] SEQ ID NO: 1039: Amino acid sequence for Lactobacillus acidophilus rhamnosidase, Uniparc reference UPI00004C6D41, Uniprot reference Q5FJ31.

[1188] SEQ ID NO: 1040: Amino acid sequence for Rhodopirellula baltica rhamnosidase, Uniparc reference UPI00001AC07D, Uniprot reference Q7UYD5.

[1189] SEQ ID NO: 1041: Amino acid sequence for Frankia inefficax rhamnosidase, Uniparc reference UPI0001BF9A6C, Uniprot reference E3IY10.

[1190] SEQ ID NO: 1042: Amino acid sequence for Streptomyces scabiei rhamnosidase, Uniparc reference UPI0001B7FF91, Uniprot reference C9Z376.

[1191] SEQ ID NO: 1043: Amino acid sequence for Flavobacterium johnsoniae rhamnosidase, Uniparc reference UPI00006E5F74, Uniprot reference A5FC22.

[1192] SEQ ID NO: 1044: Amino acid sequence for Streptomyces sp. rhamnosidase, Uniparc reference UPI00034E666D, Uniprot reference S2YWB5.

[1193] SEQ ID NO: 1045: Amino acid sequence for Acidobacterium capsulatum rhamnosidase, Uniparc reference UPI000198DF25, Uniprot reference C1F149.

[1194] SEQ ID NO: 1046: Amino acid sequence for Catenovulum agarivorans rhamnosidase, Uniparc reference UPI0003ED82D3, Uniprot reference W7QYP5.

[1195] SEQ ID NO: 1047: Amino acid sequence for Brachybacterium faecium rhamnosidase, Uniparc reference UPI0001A461B7, Uniprot reference C7MA59.

[1196] SEQ ID NO: 1048: Amino acid sequence for Klebsiella oxytoca rhamnosidase, Uniparc reference UPI000243A177, Uniprot reference A0A0J9X262.

[1197] SEQ ID NO: 1049: Amino acid sequence for Chitinophaga pinensis rhamnosidase, Uniparc reference UPI0001B24769, Uniprot reference C7PA70.

[1198] SEQ ID NO: 1050: Amino acid sequence for Streptomyces bottropensis rhamnosidase, Uniparc reference UPI0002BCAF6B, Uniprot reference M3FYL9.

[1199] SEQ ID NO: 1051: Amino acid sequence for Subdoligranulum variabile rhamnosidase, Uniparc reference UPI0001966B28, Uniprot reference D1PKC7.

[1200] SEQ ID NO: 1052: Amino acid sequence for Microbacterium testaceum rhamnosidase, Uniparc reference UPI0001F8A51D, Uniprot reference E8NDD8.

[1201] SEQ ID NO: 1053: Amino acid sequence for Solibacter usitatus rhamnosidase, Uniparc reference UPI000053767B, Uniprot reference Q01V09.

[1202] SEQ ID NO: 1054: Amino acid sequence for Streptosporangium roseum rhamnosidase, Uniparc reference UPI0001A3EFE6, Uniprot reference D2B240.

[1203] SEQ ID NO: 1055: Amino acid sequence for alpha proteobacterium rhamnosidase, Uniparc reference UPI0006CE1E82, Uniprot reference A0A0N1BME3.

[1204] SEQ ID NO: 1056: Amino acid sequence for Solitalea canadensis rhamnosidase, Uniparc reference UPI000247229E, Uniprot reference H8KPI7.

[1205] SEQ ID NO: 1057: Amino acid sequence for Parabacteroides goldsteinii rhamnosidase, Uniparc reference UPI0002CB9583, Uniprot reference S0GSF0.

[1206] SEQ ID NO: 1058: Amino acid sequence for Cyclobacterium marinum rhamnosidase, Uniparc reference UPI00021B9B33, Uniprot reference G0J630.

[1207] SEQ ID NO: 1059: Amino acid sequence for Solibacter usitatus rhamnosidase, Uniparc reference UPI0000533669, Uniprot reference Q01TX2.

[1208] SEQ ID NO: 1060: Amino acid sequence for Lunatimonas lonarensis rhamnosidase, Uniparc reference UPI00032D8F6D, Uniprot reference R7ZW70.

[1209] SEQ ID NO: 1061: Amino acid sequence for Rhizobium leguminosarum rhamnosidase, Uniparc reference UPI0000D713F2, Uniprot reference Q1M7P3.

[1210] SEQ ID NO: 1062: Amino acid sequence for Streptosporangium roseum rhamnosidase, Uniparc reference UPI0001A3EBEB, Uniprot reference D2AYU9.

[1211] SEQ ID NO: 1063: Amino acid sequence for Parabacteroides distasonis rhamnosidase, Uniparc reference UPI000156F115, Uniprot reference A6LBL4.

[1212] SEQ ID NO: 1064: Amino acid sequence for Lachnospiraceae bacterium rhamnosidase, Uniparc reference UPI0003375A10, Uniprot reference R9K6L6.

[1213] SEQ ID NO: 1065: Amino acid sequence for Chitinophaga pinensis rhamnosidase, Uniparc reference UPI0001A2F0FA, Uniprot reference C7P9Y8.

[1214] SEQ ID NO: 1066: Amino acid sequence for Caulobacter segnis rhamnosidase, Uniparc reference UPI0001BC0C15, Uniprot reference D5VGC3.

[1215] SEQ ID NO: 1067: Amino acid sequence for Pedobacter heparinus rhamnosidase, Uniparc reference UPI00019EF6E1, Uniprot reference C6Y145.

[1216] SEQ ID NO: 1068: Amino acid sequence for Pedobacter heparinus rhamnosidase, Uniparc reference UPI00019EE1A9, Uniprot reference C6Y2X3.

[1217] SEQ ID NO: 1069: Amino acid sequence for Deltaproteobacteria bacterium rhamnosidase, Uniparc reference UPI000C8D4928, Uniprot reference A0A2D5SK32.

[1218] SEQ ID NO: 1070: Amino acid sequence for Thermobaculum terrenum rhamnosidase, Uniparc reference UPI00019BFDCE, Uniprot reference D1CHL4.

[1219] SEQ ID NO: 1071: Amino acid sequence for Opitutus terrae rhamnosidase, Uniparc reference UPI000172B62A, Uniprot reference B1ZY35.

[1220] SEQ ID NO: 1072: Amino acid sequence for Kribbella flavida rhamnosidase, Uniparc reference UPI00019BFABB, Uniprot reference D2PXQ4.

[1221] SEQ ID NO: 1073: Amino acid sequence for Streptomyces scabiei rhamnosidase, Uniparc reference UPI0001B80091, Uniprot reference C9Z391.

[1222] SEQ ID NO: 1074: Amino acid sequence for Actinoplanes sp. rhamnosidase, Uniparc reference UPI00023EC5D1, Uniprot reference G8S540.

[1223] SEQ ID NO: 1075: Amino acid sequence for Asticcacaulis sp. rhamnosidase, Uniparc reference UPI0003C3CD2B, Uniprot reference V4NSJ1.

[1224] SEQ ID NO: 1076: Amino acid sequence for Kribbella flavida rhamnosidase, Uniparc reference UPI00019BF65D, Uniprot reference D2PT74.

[1225] SEQ ID NO: 1077: Amino acid sequence for Bacillus sp. rhamnosidase, Uniparc reference UPI00000BC760, Uniprot reference Q93RE7.

[1226] SEQ ID NO: 1078: Amino acid sequence for Flavobacterium johnsoniae rhamnosidase, Uniparc reference UPI00006E5FAB, Uniprot reference A5FCG3.

[1227] SEQ ID NO: 1079: Amino acid sequence for Lunatimonas lonarensis rhamnosidase, Uniparc reference UPI00032EEB9C, Uniprot reference R7ZS84.

[1228] SEQ ID NO: 1080: Amino acid sequence for Eisenbergiella massiliensis rhamnosidase, Uniparc reference UPI0004B2D794, Uniprot reference A0A3E3IGR6.

[1229] SEQ ID NO: 1081: Amino acid sequence for Catenovulum agarivorans rhamnosidase, Uniparc reference UPI0003ED7515, Uniprot reference W7QF25.

[1230] SEQ ID NO: 1083: Codon optimised DNA encoding 6×His Rhodothermus marinus rhamnosidase, Uniparc reference UPI0001A31108, Uniprot reference D0MFR0.

[1231] SEQ ID NO: 1084: Codon optimised DNA encoding 6×His Streptomyces bingchenggensis rhamnosidase, Uniparc reference UPI0001D90BFE, Uniprot reference D7C463.

[1232] SEQ ID NO: 1085: Codon optimised DNA encoding 6×His Spirosoma linguale rhamnosidase Uniparc reference UPI0001A3AEAC, Uniprot reference D2QL60.

[1233] SEQ ID NO: 1086: Codon optimised DNA encoding 6×His Roseburia intestinalis rhamnosidase, Uniparc reference UPI0001CD6D48, Uniprot reference D4L2K8.

[1234] SEQ ID NO: 1087: Codon optimised DNA encoding 6×His Draconibacterium orientale rhamnosidase, Uniparc reference UPI000442EF22, Uniprot reference X5DG83.

[1235] SEQ ID NO: 1088: Codon optimised DNA encoding 6×His Catenulispora acidiphila rhamnosidase, Uniparc reference UPI00019E052B, Uniprot reference C7QC24.

[1236] SEQ ID NO: 1089: Codon optimised DNA encoding 6×His Bacteroides thetaiotaomicron rhamnosidase, Uniparc reference UPI000005ADE1, Uniprot reference Q8A916.

[1237] SEQ ID NO: 1090: Codon optimised DNA encoding 6×His Opitutus terrae rhamnosidase, Uniparc reference UPI000172B2E3, Uniprot reference B1ZRE4.

[1238] SEQ ID NO: 1091: Codon optimised DNA encoding 6×His Lachnoclostridium phytofermentans rhamnosidase, Uniparc reference UPI00015FE0BE, Uniprot reference A9KJP8.

[1239] SEQ ID NO: 1092: Codon optimised DNA encoding 6×His Rhodanobacter denitrificans rhamnosidase, Uniparc reference UPI00022DA3ED, Uniprot reference M4NH01.

[1240] SEQ ID NO: 1093: Codon optimised DNA encoding 6×His Prevotella ruminicola rhamnosidase, Uniparc reference UPI0001D07633, Uniprot reference D5ETD9.

[1241] SEQ ID NO: 1094: Codon optimised DNA encoding 6×His Aspergillus terreus rhamnosidase Uniparc reference UPI000259E388, Uniprot reference 10AZ41.

[1242] SEQ ID NO: 1095: Codon optimised DNA encoding 6×His Brachybacterium faecium rhamnosidase, Uniparc reference UPI0001A461B6, Uniprot reference C7MA58.

[1243] SEQ ID NO: 1096: Codon optimised DNA encoding 6×His Flavobacterium johnsoniae rhamnosidase, Uniparc reference UPI00006E4A0D, Uniprot reference A5FCH3.

[1244] SEQ ID NO: 1097: Codon optimised DNA encoding 6×His Rahnella aquatilis rhamnosidase, Uniparc reference UPI000245C507, Uniprot reference H2IYR2.

[1245] SEQ ID NO: 1098: Codon optimised DNA encoding 6×His Bifidobacterium moukalabense rhamnosidase, Uniparc reference UPI0003E4E711, Uniprot reference W4N6H0.

[1246] SEQ ID NO: 1099: Codon optimised DNA encoding 6×His Enterococcus casseliflavus rhamnosidase, Uniparc reference UPI000353BD9F, Uniprot reference S4BBS9.

[1247] SEQ ID NO: 1100: Codon optimised DNA encoding 6×His Geobacillus sp. rhamnosidase, Uniparc reference UPI0001789COD, Uniprot reference D3EED1.

[1248] SEQ ID NO: 1101: Codon optimised DNA encoding 6×His Modestobacter marinus rhamnosidase, Uniparc reference UPI000260A2FE, Uniprot reference I4EYD9.

[1249] SEQ ID NO: 1102: Codon optimised DNA encoding 6×His Pedobacter heparinus rhamnosidase, Uniparc reference UPI00019EF0EE, Uniprot reference C6XYM6.

[1250] SEQ ID NO: 1103: Codon optimised DNA encoding 6×His Dyadobacter fermentans rhamnosidase, Uniparc reference UPI00019B5915, Uniprot reference C6VZL3.

[1251] SEQ ID NO: 1104: Codon optimised DNA encoding 6×His Paenibacillus mucilaginosus rhamnosidase, Uniparc reference UPI000212C156, Uniprot reference F8FQQ3.

[1252] SEQ ID NO: 1105: Codon optimised DNA encoding 6×His Paenibacillus sp. rhamnosidase, Uniparc reference UPI0003E2544D, Uniprot reference W4D866.

[1253] SEQ ID NO: 1106: Codon optimised DNA encoding 6×His Bacteroides thetaiotaomicron rhamnosidase, Uniparc reference UPI000005ADD2, Uniprot reference A0A0P0FM19.

[1254] SEQ ID NO: 1107: Codon optimised DNA encoding 6×His Chloroflexus aurantiacus rhamnosidase, Uniparc reference UPI00005BA60B, Uniprot reference A9WDK5.

[1255] SEQ ID NO: 1108: Codon optimised DNA encoding 6×His Thermoclostridium stercorarium rhamnosidase, Uniparc reference UPI00000B098C, Uniprot reference Q9S3L0.

[1256] SEQ ID NO: 1109: Codon optimised DNA encoding 6×His Bifidobacterium moukalabense rhamnosidase, Uniparc reference UPI0003E5C314, Uniprot reference W4N612.

[1257] SEQ ID NO: 1110: Codon optimised DNA encoding 6×His Olsenella profusa rhamnosidase, Uniparc reference UPI0003AE032C, Uniprot reference U2USP4.

[1258] SEQ ID NO: 1111: Codon optimised DNA encoding 6×His Kribbella flavida rhamnosidase, Uniparc reference UPI00019BDB13, Uniprot reference D2PMT5.

[1259] SEQ ID NO: 1112: Codon optimised DNA encoding 6×His Caulobacter vibrioides rhamnosidase, Uniparc reference UPI00000C7226, Uniprot reference Q9A9K2.

[1260] SEQ ID NO: 1113: Codon optimised DNA encoding 6×His Bacteroides thetaiotaomicron rhamnosidase, Uniparc reference UPI000005BA09, Uniprot reference Q8A076.

[1261] SEQ ID NO: 1114: Codon optimised DNA encoding 6×His Rhodonellum psychrophilum rhamnosidase, Uniparc reference UPI0003745394, Uniprot reference U5BUY4.

[1262] SEQ ID NO: 1115: Codon optimised DNA encoding 6×His Paenibacillus sp. rhamnosidase, Uniparc reference UPI0004F6D660, Uniprot reference A0A089M3T2.

[1263] SEQ ID NO: 1116: Codon optimised DNA encoding 6×His Catenovulum agarivorans rhamnosidase, Uniparc reference UPI0003ED860D, Uniprot reference W7QMH5.

[1264] SEQ ID NO: 1117: Codon optimised DNA encoding 6×His Zobellia galactanivorans rhamnosidase, Uniparc reference UPI000217D8B1, Uniprot reference G0L382.

[1265] SEQ ID NO: 1118: Codon optimised DNA encoding 6×His Bacteroides thetaiotaomicron rhamnosidase, Uniparc reference UPI000005B845, Uniprot reference Q8A1H5.

[1266] SEQ ID NO: 1119: Codon optimised DNA encoding 6×His Bacteroides xylanisolvens rhamnosidase, Uniparc reference UPI0001CD02E9, Uniprot reference D6CYE5.

[1267] SEQ ID NO: 1120: Codon optimised DNA encoding 6×His Pseudarthrobacter chlorophenolicus rhamnosidase, Uniparc reference UPI00018E07C6, Uniprot reference B8HAH3.

[1268] SEQ ID NO: 1121: Codon optimised DNA encoding 6×His Dictyoglomus thermophilum rhamnosidase, Uniparc reference UPI0001815896, Uniprot reference B5YC64.

[1269] SEQ ID NO: 1122: Codon optimised DNA encoding 6×His Formosa agariphila rhamnosidase, Uniparc reference UPI00039231C1, Uniprot reference T2KPL4.

[1270] SEQ ID NO: 1123: Codon optimised DNA encoding 6×His Rhodococcus jostii rhamnosidase, Uniparc reference UPI0000DBA6EB, Uniprot reference Q0S9T4.

[1271] SEQ ID NO: 1124: Codon optimised DNA encoding 6×His Lactobacillus crispatus rhamnosidase, Uniparc reference UPI0001D10896, Uniprot reference D5GZ45.

[1272] SEQ ID NO: 1125: Codon optimised DNA encoding 6×His Pedobacter heparinus rhamnosidase, Uniparc reference UPI0001B17DE6, Uniprot reference C6XVU2.

[1273] SEQ ID NO: 1126: Codon optimised DNA encoding 6×His Spirosoma linguale rhamnosidase, Uniparc reference UPI0001A3C989, Uniprot reference D2QUA5.

[1274] SEQ ID NO: 1127: Codon optimised DNA encoding 6×His Pedobacter heparinus rhamnosidase, Uniparc reference UPI00019EE3EB, Uniprot reference C6XU05.

[1275] SEQ ID NO: 1128: Codon optimised DNA encoding 6×His Paenibacillus mucilaginosus rhamnosidase, Uniparc reference UPI0003432C11, Uniprot reference R9ULQ4.

[1276] SEQ ID NO: 1129: Codon optimised DNA encoding 6×His Caulobacter segnis rhamnosidase, Uniparc reference UPI0001BC0C05, Uniprot reference D5VGD9.

[1277] SEQ ID NO: 1130: Codon optimised DNA encoding 6×His Bacteroides cellulosilyticus rhamnosidase, Uniparc reference UPI0001969377, Uniprot reference E2N9B1.

[1278] SEQ ID NO: 1131: Codon optimised DNA encoding 6×His Pedobacter heparinus rhamnosidase, Uniparc reference UPI0001B17C60, Uniprot reference C6Y153.

[1279] SEQ ID NO: 1132: Codon optimised DNA encoding 6×His Formosa agariphila rhamnosidase, Uniparc reference UPI000571C0C2, Uniprot reference T2KNB2.

[1280] SEQ ID NO: 1133: Codon optimised DNA encoding 6×His Lactobacillus acidophilus rhamnosidase, Uniparc reference UPI00004C6D41, Uniprot reference Q5FJ31.

[1281] SEQ ID NO: 1134: Codon optimised DNA encoding 6×His Rhodopirellula baltica rhamnosidase, Uniparc reference UPI00001AC07D, Uniprot reference Q7UYD5.

[1282] SEQ ID NO: 1135: Codon optimised DNA encoding 6×His Frankia inefficax rhamnosidase, Uniparc reference UPI0001BF9A6C, Uniprot reference E3IY10.

[1283] SEQ ID NO: 1136: Codon optimised DNA encoding 6×His Streptomyces scabiei rhamnosidase, Uniparc reference UPI0001B7FF91, Uniprot reference C9Z376.

[1284] SEQ ID NO: 1137: Codon optimised DNA encoding 6×His Flavobacterium johnsoniae rhamnosidase, Uniparc reference UPI00006E5F74, Uniprot reference A5FC22.

[1285] SEQ ID NO: 1138: Codon optimised DNA encoding 6×His Streptomyces sp. rhamnosidase, Uniparc reference UPI00034E666D, Uniprot reference S2YWB5.

[1286] SEQ ID NO: 1139: Codon optimised DNA encoding 6×His Acidobacterium capsulatum rhamnosidase, Uniparc reference UPI000198DF25, Uniprot reference C1F149.

[1287] SEQ ID NO: 1140: Codon optimised DNA encoding 6×His Catenovulum agarivorans rhamnosidase, Uniparc reference UPI0003ED82D3, Uniprot reference W7QYP5.

[1288] SEQ ID NO: 1141: Codon optimised DNA encoding 6×His Brachybacterium faecium rhamnosidase, Uniparc reference UPI0001A461B7, Uniprot reference C7MA59.

[1289] SEQ ID NO: 1142: Codon optimised DNA encoding 6×His Klebsiella oxytoca rhamnosidase, Uniparc reference UPI000243A177, Uniprot reference A0A0J9X262.

[1290] SEQ ID NO: 1143: Codon optimised DNA encoding 6×His Chitinophaga pinensis rhamnosidase, Uniparc reference UPI0001B24769, Uniprot reference C7PA70.

[1291] SEQ ID NO: 1144: Codon optimised DNA encoding 6×His Streptomyces bottropensis rhamnosidase, Uniparc reference UPI0002BCAF6B, Uniprot reference M3FYL9.

[1292] SEQ ID NO: 1145: Codon optimised DNA encoding 6×His Subdoligranulum variabile rhamnosidase, Uniparc reference UPI0001966B28, Uniprot reference D1PKC7.

[1293] SEQ ID NO: 1146: Codon optimised DNA encoding 6×His Microbacterium testaceum rhamnosidase, Uniparc reference UPI0001F8A51D, Uniprot reference E8NDD8.

[1294] SEQ ID NO: 1147: Codon optimised DNA encoding 6×His Solibacter usitatus rhamnosidase, Uniparc reference UPI000053767B, Uniprot reference Q01V09.

[1295] SEQ ID NO: 1148: Codon optimised DNA encoding 6×His Streptosporangium roseum rhamnosidase, Uniparc reference UPI0001A3EFE6, Uniprot reference D2B240.

[1296] SEQ ID NO: 1149: Codon optimised DNA encoding 6×His alpha proteobacterium rhamnosidase, Uniparc reference UPI0006CE1E82, Uniprot reference A0A0N1 BME3.

[1297] SEQ ID NO: 1150: Codon optimised DNA encoding 6×His Solitalea canadensis rhamnosidase, Uniparc reference UPI000247229E, Uniprot reference H8KP17.

[1298] SEQ ID NO: 1151: Codon optimised DNA encoding 6×His Parabacteroides goldsteinii rhamnosidase, Uniparc reference UPI0002CB9583, Uniprot reference S0GSF0.

[1299] SEQ ID NO: 1152: Codon optimised DNA encoding 6×His Cyclobacterium marinum rhamnosidase, Uniparc reference UPI00021B9B33, Uniprot reference G0J630.

[1300] SEQ ID NO: 1153: Codon optimised DNA encoding 6×His Solibacter usitatus rhamnosidase, Uniparc reference UPI0000533669, Uniprot reference Q01TX2.

[1301] SEQ ID NO: 1154: Codon optimised DNA encoding 6×His Lunatimonas lonarensis rhamnosidase, Uniparc reference UPI00032D8F6D, Uniprot reference R7ZW70.

[1302] SEQ ID NO: 1155: Codon optimised DNA encoding 6×His Rhizobium leguminosarum rhamnosidase, Uniparc reference UPI0000D713F2, Uniprot reference Q1M7P3.

[1303] SEQ ID NO: 1156: Codon optimised DNA encoding 6×His Streptosporangium roseum rhamnosidase, Uniparc reference UPI0001A3EBEB, Uniprot reference D2AYU9.

[1304] SEQ ID NO: 1157: Codon optimised DNA encoding 6×His Parabacteroides distasonis rhamnosidase, Uniparc reference UPI000156F115, Uniprot reference A6LBL4.

[1305] SEQ ID NO: 1158: Codon optimised DNA encoding 6×His Lachnospiraceae bacterium rhamnosidase, Uniparc reference UPI0003375A10, Uniprot reference R9K6L6.

[1306] SEQ ID NO: 1159: Codon optimised DNA encoding 6×His Chitinophaga pinensis rhamnosidase, Uniparc reference UPI0001A2F0FA, Uniprot reference C7P9Y8.

[1307] SEQ ID NO: 1160: Codon optimised DNA encoding 6×His Caulobacter segnis rhamnosidase, Uniparc reference UPI0001 BC0C15, Uniprot reference D5VGC3.

[1308] SEQ ID NO: 1161: Codon optimised DNA encoding 6×His Pedobacter heparinus rhamnosidase, Uniparc reference UPI00019EF6E1, Uniprot reference C6Y145.

[1309] SEQ ID NO: 1162: Codon optimised DNA encoding 6×His Pedobacter heparinus rhamnosidase, Uniparc reference UPI00019EE1A9, Uniprot reference C6Y2X3.

[1310] SEQ ID NO: 1163: Codon optimised DNA encoding 6×His Deltaproteobacteria bacterium rhamnosidase, Uniparc reference UPI000C8D4928, Uniprot reference A0A2D5SK32.

[1311] SEQ ID NO: 1164: Codon optimised DNA encoding 6×His Thermobaculum terrenum rhamnosidase, Uniparc reference UPI00019BFDCE, Uniprot reference D1CHL4.

[1312] SEQ ID NO: 1165: Codon optimised DNA encoding 6×His Opitutus terrae rhamnosidase, Uniparc reference UPI000172B62A, Uniprot reference B1ZY35.

[1313] SEQ ID NO: 1166: Codon optimised DNA encoding 6×His Kribbella flavida rhamnosidase, Uniparc reference UPI00019BFABB, Uniprot reference D2PXQ4.

[1314] SEQ ID NO: 1167: Codon optimised DNA encoding 6×His Streptomyces scabiei rhamnosidase, Uniparc reference UPI0001B80091, Uniprot reference C9Z391.

[1315] SEQ ID NO: 1168: Codon optimised DNA encoding 6×His Actinoplanes sp. rhamnosidase, Uniparc reference UPI00023EC5D1, Uniprot reference G8S540.

[1316] SEQ ID NO: 1169: Codon optimised DNA encoding 6×His Asticcacaulis sp. rhamnosidase, Uniparc reference UPI0003C3CD2B, Uniprot reference V4NSJ1.

[1317] SEQ ID NO: 1170: Codon optimised DNA encoding 6×His Kribbella flavida rhamnosidase, Uniparc reference UPI00019BF65D, Uniprot reference D2PT74.

[1318] SEQ ID NO: 1171: Codon optimised DNA encoding 6×His Bacillus sp. rhamnosidase, Uniparc reference UPI00000BC760, Uniprot reference Q93RE7.

[1319] SEQ ID NO: 1172: Codon optimised DNA encoding 6×His Flavobacterium johnsoniae rhamnosidase, Uniparc reference UPI00006E5FAB, Uniprot reference A5FCG3.

[1320] SEQ ID NO: 1173: Codon optimised DNA encoding 6×His Lunatimonas lonarensis rhamnosidase, Uniparc reference UPI00032EEB9C, Uniprot reference R7ZS84.

[1321] SEQ ID NO: 1174: Codon optimised DNA encoding 6×His Eisenbergiella massiliensis rhamnosidase, Uniparc reference UPI0004B2D794, Uniprot reference A0A3E3IGR6.

[1322] SEQ ID NO: 1175: Codon optimised DNA encoding 6×His Catenovulum agarivorans rhamnosidase, Uniparc reference UPI0003ED7515, Uniprot reference W7QF25.

[1323] SEQ ID NO: 1176: Codon optimised DNA encoding 6×His Streptomyces avermitilis rhamnosidase, Uniparc reference UPI0000184198, Uniprot reference Q82PP4.

[1324] SEQ ID NO: 1177: N-terminal His-tag / linker

[1325] SEQ ID NO: 1178: C-terminal linker / His tag

[1326] SEQ ID NO: 1179: Amino acid sequence for Variant G1.

[1327] SEQ ID NO: 1180: Amino acid sequence for Variant G2.

[1328] SEQ ID NO: 1181: Amino acid sequence for Variant G3.

[1329] SEQ ID NO: 1182: Amino acid sequence for Variant G4.

[1330] SEQ ID NO: 1183: Amino acid sequence for Variant G5.

[1331] SEQ ID NO: 1184: Codon optimised DNA encoding N-terminal 6×His Variant G1.

[1332] SEQ ID NO: 1185: Codon optimised DNA encoding N-terminal 6×His Variant G2.

[1333] SEQ ID NO: 1186: Codon optimised DNA encoding N-terminal 6×His Variant G3.

[1334] SEQ ID NO: 1187: Codon optimised DNA encoding N-terminal 6×His Variant G4.

[1335] SEQ ID NO: 1188: Codon optimised DNA encoding N-terminal 6×His Variant G5.

[1336] SEQ ID NO: 1189: Amino acid sequence for Variant R1.

[1337] SEQ ID NO: 1190: Amino acid sequence for Variant R2.

[1338] SEQ ID NO: 1191: Amino acid sequence for Variant R3.

[1339] SEQ ID NO: 1192: Amino acid sequence for Variant R4.

[1340] SEQ ID NO: 1193: Amino acid sequence for Variant R5.

[1341] SEQ ID NO: 1194: Codon optimised DNA encoding C-terminal 6×His Variant R1.

[1342] SEQ ID NO: 1195: Codon optimised DNA encoding C-terminal 6×His Variant R2.

[1343] SEQ ID NO: 1196: Codon optimised DNA encoding C-terminal 6×His Variant R3.

[1344] SEQ ID NO: 1197: Codon optimised DNA encoding C-terminal 6×His Variant R4.

[1345] SEQ ID NO: 1198: Codon optimised DNA encoding C-terminal 6×His Variant R5.DETAILED DESCRIPTION OF THE INVENTION

[1346] As mentioned previously, saponins are steroid or terpenoid glycosides which have a broad range of uses. Current approaches to obtaining certain saponins, such as from plant cell culture, in suitable quantities and of suitable purities are limiting. The present inventors have surprisingly found that enzymatic modification of saponins synthesized and produced by plant cell culture can facilitate improved availability of saponins of interest and / or facilitate removal of undesired saponin components. The present invention therefore provides methods for the enzymatic modification of saponins synthesized and produced by plant cell culture, products made by such methods, uses of said products and associated aspects. In methods of the invention, a starting saponin (i.e. a saponin to be modified by an enzyme) is converted into a product saponin (i.e. the saponin resulting from enzymatic modification of the starting saponin).

[1347] Engineered glucosidase polypeptides disclosed herein may be used in methods for the enzymatic modification of saponins of the present invention.

[1348] Engineered rhamnosidase polypeptides disclosed herein may be used in methods for the enzymatic modification of saponins of the present invention.Objectives

[1349] The methods of the present invention can be applied to achieve a plurality of objectives, such as: (i) improving the yield of saponins of interest obtainable from a given starting material; (ii) broadening the range of starting materials suitable for obtaining saponins of interest; and / or (iii) convenient removal of undesired saponins from saponins of interest. In the sense of the present invention, the term ‘starting material’ designates materials deriving from a plant cell culture.

[1350] Where supply is constrained for a starting material from which a saponin of interest is isolated, achieving the maximum yield of the saponin of interest is clearly important. Independently of the efficiency of extraction and separation processes which would generally be adapted for optimal isolation of existing saponin of interest, the present invention may be applied to increase the amount of a saponin of interest which may be obtained from a given starting material. Enzymatic modification of other saponins present in the starting material to form a saponin of interest can increase the amount of the saponin of interest which may be obtained, by enriching the starting material with the saponin of interest.

[1351] Saponins may be obtained from a broad range of sources. The presence of specific saponins and their levels in plant materials may depend on a range of factors such as a plant species, tissue, age, season, environmental conditions and the like. Variation may also be observed between individual plants (such as trees) of the same species (see, for example, WO2018057031). Variation in the level of specific saponins, and the respective proportion thereof, may be similarly observed from one plant cell culture to another. The burden associated with extraction and / or isolation of a saponin of interest may mean that certain potential sources, such as in vitro plant cell culture, of the saponin of interest are not commercially viable, due to the saponin of interest being present at relatively low levels. Enzymatic modification of other saponins present to form a saponin of interest can therefore expand the range of viable sources for obtaining the saponin of interest.

[1352] It is well understood that different saponins may have different activity profiles—both positive / desired activities and negative / undesired activities. Some uses of saponins require a high degree of purification and separating a saponin of interest from other saponins, particularly those of similar structure or physical properties, can be burdensome. Enzymatic modification of such other saponins may alter their physical properties and may thereby facilitate separation from a saponin of interest. Other uses of saponins may not require a high degree of purity per se, nevertheless it may still be desirable to remove or reduce the amount of a particular saponin component (or components) within a saponin mixture without burdensome chromatographic methods. Enzymatic modification can facilitate removal or reduction in the level of a particular saponin component within a saponin mixture without the need for chromatographic means.Saponins

[1353] The methods of the present invention require a starting saponin (i.e. a saponin which is intended to be enzymatically modified). The starting saponin may be a naturally occurring saponin (i.e. a steroid or terpenoid glycoside found in nature), or an artificially created saponin (i.e. a steroid or terpenoid glycoside not found in nature).

[1354] In some embodiments, the starting saponin is a steroid glycoside. In other embodiments, the starting saponin is a terpenoid glycoside, especially a triterpenoid glycoside.

[1355] Naturally occurring starting saponins may be saponins synthesized by plant cells cultured in vitro that originate from plants that produce saponins, such as the plants described below.

[1356] Naturally occurring starting saponins include those obtainable from, such as obtained from, an in vitro plant cell culture originating from the genera Gypsophilia, Saponaria, such as the Saponaria vaccaria species, or Saponaria officinalis species, or Quillaja (Bomford, 1992). Especially of interest are starting saponins obtainable, such a obtained, from an in vitro plant cell culture originating from Quillaja species. Particular starting saponins of interest include those obtained from an in vitro plant cell culture originating from Quillaja brasiliensis or Quillaja saponaria. In one embodiment, the starting saponin is obtained from an in vitro plant cell culture originating from Quillaja saponaria. In one embodiment, the starting saponin is obtained from an in vitro plant cell culture originating from Quillaja brasiliensis.

[1357] In certain embodiments, the starting saponin is a quillaic acid glycoside. In certain embodiments, the starting saponin is a phytolaccinic acid glycoside. In certain embodiments, the starting saponin is an echinocystic acid glycoside. In certain embodiments, the starting saponin is a 22-beta-hydroxylated quillaic acid glycoside. In certain embodiments, the starting saponin is a gypsogenin glycoside.

[1358] Analysis of water / methanol extracts of Quillaja saponaria bark by liquid chromatography / mass spectrometry has revealed over 100 saponins (Nyberg, 2000; Nyberg, 2003; Kite, 2004). Quillaja brasiliensis extracts have also been described, with many saponin components in Quillaja brasiliensis extracts corresponding to saponins found in Quillaja saponaria extracts. (Wallace, 2017; Wallace, 2019).

[1359] The following text describes particular quillaic acid-derived starting and product saponins which are grouped by ‘family’. Each family has one or more common structural features which characterise the family relative to other families. Individual components within each family also display certain structural features which characterise the component relative to other components of the family, including: xylose or rhamnose chemotype—the presence of a xylose or rhamnose residue in the C3 saccharide; A or B isomers—A having the acyl chain linked through the 4-position of the D-fucose, B having the acyl chain linked through the 3-position of the D-fucose; V1 and V2—the presence of a terminal apiose or xylose residue, respectively, in the C28 saccharide (in other components of a family this terminal residue may also be absent). The text focuses on components which typically have a significant presence in Quillaja saponaria aqueous extracts, such as bark extracts thereof, but it will be appreciated that (i) other components of a family also exist and (ii) the proportions of different components of a family may vary both between families and between different saponin sources (Kite, 2004). The specific extraction method used may also influence the proportions of different components obtained. While saponin components have been extensively studied using bark extracts of Quillaja saponaria, it is assumed that the saponin components synthesized and produced by plant cells cultured in vitro that originate from Quillaja saponaria would be the same.

[1360] A and B isomers may be separable using chromatographic techniques. However, under suitable solvent conditions these isomers will revert to equilibrium proportions (see e.g. Cleland, 1996). Xylose and rhamnose chemotypes typically elute closely. Depending on chromatographic techniques, the rhamnose chemotype may form a minor peak closely preceding or overlapping with the main peak for the family.

[1361] Those skilled in the art will also recognise that the structures described contain ionisable groups and under appropriate circumstances may exist in dissociated forms or as salts. Structures are generally shown with the glucuronate moiety in ionised form and the indicated molecular weight is calculated directly from the ion shown (corresponding to the monoisotopic m / z observed with negative mode electrospray mass spectrometry), however, all non-dissociated, dissociated and salt forms are intended to be encompassed by the recited definitions. Salts are desirably pharmaceutically acceptable, although non-pharmaceutically acceptable salts can nevertheless be useful during manufacture of pharmaceuticals or for non-pharmaceutical uses.

[1362] Starting saponins obtainable from plant cells cultured in vitro and originating from Quillaja saponaria include:

[1363] QS-18 family components (i.e. triterpenoid glycosides having beta-O-glucopyranosylation at the C3 position of the L-rhamnose moiety of QS-21 family components), such as:

[1364] ‘QS-18 2150 A component’, being the triterpenoid glycosides identified as part of the QS-18 main peak in FIG. 2 and having a m / z of 2150 with negative mode electrospray mass spectrometry. The QS-18 2150 A component is believed to be identified in Kite 2004 as Peak 76 and corresponds to the A-isomer xylose chemotype structures B4 (apiose isomer) and B6 (xylose isomer) characterised in Nyberg 2000 and Nyberg 2003. The QS-18 2150 A component may consist of QS-18 2150 A V1 (i.e. apiose isomer): and / or QS-18 2150 A V2 (i.e. xylose isomer):‘QS-18 2018 A component’, being the triterpenoid glycosides identified as part of the QS-18 main peak in FIG. 2 and having m / z of 2018 with negative mode electrospray mass spectrometry. The QS-18 A 2018 component is believed to be identified in Kite 2004 as Peak 73 and corresponds to the A-isomer xylose chemotype structure B2 characterised in Nyberg 2000 and Nyberg 2003. The QS-18 2018 A component may consist of QS-18 2018 A:‘QS-18 2164 A component’, being the triterpenoid glycosides identified as part of the QS-18 main peak in FIG. 2 and having a m / z of 2164 with negative mode electrospray mass spectrometry. The QS-18 2164 A component is believed to be identified in Kite 2004 as Peak 74 and corresponds to the A-isomer rhamnose chemotype structures B3 (apiose isomer) and B5 (xylose isomer) characterised in Nyberg 2000 and Nyberg 2003. The QS-18 2164 A component may consist of QS-18 2164 A V1 (i.e. apiose isomer): and / or QS-18 2164 A V2 (i.e. xylose isomer):‘QS-18 2150 B component’, being triterpenoid glycosides having a m / z of 2150 with negative mode electrospray mass spectrometry. The QS-18 2150 B component corresponds to the B-isomer xylose chemotype structures B4a (apiose isomer) and B6a (xylose isomer) characterised in Nyberg 2000 and Nyberg 2003. The QS-18 2150 B component may consist of QS-18 2150 B V1 (i.e. apiose isomer): and / or QS-18 2150 B V2 (i.e. xylose isomer):‘QS-18 2018 B component’, being triterpenoid glycosides having a m / z of 2018 with negative mode electrospray mass spectrometry. The QS-18 2018 B component corresponds to the B-isomer xylose chemotype structure B2a characterised in Nyberg 2000 and Nyberg 2003. The QS-18 2018 B component may consist of QS-18 2018 B:‘QS-18 2164 B component’, being triterpenoid glycosides having a m / z of 2164 with negative mode electrospray mass spectrometry. The QS-18 2164 B corresponds to the B-isomer rhamnose chemotype structures B3a (apiose isomer) and B5a (xylose isomer). The QS-18 2164 B component may consist of QS-18 2164 B V1 (i.e. apiose isomer): and / or QS-18 2164 B V2 (i.e. xylose isomer):desglucosyl-QS-17 family components (i.e. triterpenoid glycosides having alpha-O-rhamnosylation at the C2 position of the arabinofuranose moiety of QS-21 family components but lacking the glycosylation of QS-17 family components), such as:‘desglucosyl-QS-17 2134 A component’, being triterpenoid glycosides having a m / z of 2134 with negative mode electrospray mass spectrometry. The desglucosyl-QS-17 2134 A component is believed to be identified in Kite 2004 as Peak 75 and corresponds to A-isomers of the xylose chemotype. The desglucosyl-QS-17 2134 A component may consist of desglucosyl-QS-17 2134 A V1 (i.e. apiose isomer): and / or desglucosyl-QS-17 2134 A V2 (i.e. xylose isomer):‘desglucosyl-QS-17 2002 A component’, being triterpenoid glycosides having a m / z of 2002 with negative mode electrospray mass spectrometry. The‘desglucosyl-QS-17 2002 A component’ corresponds to the A-isomer xylose chemotype and may consist of desglucosyl-QS-17 2002 A:‘desglucosyl-QS-17 2148 A component’, being triterpenoid glycosides having a m / z of 2148 with negative mode electrospray mass spectrometry. The desglucosyl-QS-17 2148 A component is believed to be identified in Kite 2004 as Peaks 70 and 72 and corresponds to the A-isomer rhamnose chemotype. The desglucosyl-QS-17 2148 A component may consist of desglucosyl-QS-17 2148 A V1 (i.e. apiose isomer): and / or desglucosyl-QS-17 2148 A V2 (i.e. xylose isomer):‘desglucosyl-QS-17 2134 B component’, being triterpenoid glycosides having a m / z of 2134 with negative mode electrospray mass spectrometry. The desglucosyl-QS-17 2134 B component is believed to be identified in Kite 2004 as Peak 67 and corresponds to B-isomers of the xylose chemotype. The desglucosyl-QS-17 2134 B component may consist of desglucosyl-QS-17 2134 B V1 (i.e. apiose isomer): and / or desglucosyl-QS-17 2134 B V2 (i.e. xylose isomer):‘desglucosyl-QS-17 2002 B component’, being triterpenoid glycosides having a m / z of 2002 with negative mode electrospray mass spectrometry. The desglucosyl-QS-17 2002 B component corresponds to the B-isomer of the xylose chemotype and may consist of desglucosyl-QS-17 2002 B:‘desglucosyl-QS-17 2148 B component’, being triterpenoid glycosides having a m / z of 2148 with negative mode electrospray mass spectrometry. The desglucosyl-QS-17 2148 B component is believed to be identified in Kite 2004 as Peak 65 and corresponds to B-isomers of the rhamnose chemotype. The desglucosyl-QS-17 2148 B component may consist of desglucosyl-QS-17 2148 B V1 (i.e. apiose isomer): and / or desglucosyl-QS-17 2148 B V2 (i.e. xylose isomer):QS-17 family components (i.e. triterpenoid glycosides having beta-O-glucopyranosylation at the C3 position of the L-rhamnose moiety and alpha-O-rhamnosylation at the C2 position of the arabinofuranose moiety of QS-21 family components), such as:‘QS-17 2296 A component’, being the triterpenoid glycosides identified as part of the QS-17 main peak in FIG. 2 and having a m / z of 2296 with negative mode electrospray mass spectrometry. The QS-17 2296 A component is believed to be identified in Kite 2004 as Peak 59 and corresponds to the A-isomers of xylose chemotype structure QS-III. The QS-17 2296 A component may consist of QS-17 2296 A V1 (i.e. apiose isomer): and / or QS-17 2296 A V2 (i.e. xylose isomer):‘QS-17 2164 A component’, being the triterpenoid glycosides identified as part of the QS-17 main peak in FIG. 2 and having a m / z of 2164 with negative mode electrospray mass spectrometry. The QS-17 2164 A component is believed to be identified in Kite 2004 as Peak 58 and corresponds to the A-isomer xylose chemotype. The QS-17 2164 A component may consist of QS-17 2164 A:‘QS-17 2310 A component’, being the triterpenoid glycosides identified as part of the QS-17 main peak in FIG. 2 and having a m / z of 2310 with negative mode electrospray mass spectrometry. The QS-17 2310 A component is believed to be identified in Kite 2004 as Peak 57 and corresponds to A-isomers of the rhamnose chemotype. The QS-17 2310 A component may consist of QS-17 2310 A V1 (i.e. apiose isomer): and / or QS-17 2310 A V2 (i.e. xylose isomer):‘QS-17 2296 B component’, being triterpenoid glycosides having a m / z of 2296 with negative mode electrospray mass spectrometry. The QS-17 2296 B component corresponds to the B-isomers of xylose chemotype structure QS-III in Kite 2004. The QS-17 2296 B component may consist of QS-17 2296 B V1 (i.e. apiose isomer): and / or QS-17 2296 B V2 (i.e. xylose isomer):‘QS-17 2164 B component’, being triterpenoid glycosides having a m / z of 2164 with negative mode electrospray mass spectrometry. The QS-17 2164 B component corresponds to the B-isomer xylose chemotype. The QS-17 2164 B component and may consist of QS-17 2164 B:‘QS-17 2310 B component’, being triterpenoid having a m / z of 2310 with negative mode electrospray mass spectrometry. The QS-17 2310 B component corresponds to B-isomers of the rhamnose chemotype. The QS-17 2310 B component may consist of QS-17 2310 B V1 (i.e. apiose isomer): and / or QS-17 2310 B V2 (i.e. xylose isomer);desarabinofuranosyl-QS-18 family components (i.e. triterpenoid glycosides having beta-O-glucopyranosylation at the C3 position of the L-rhamnose moiety and but lacking the arabinofuranose moiety of QS-21 family components). The desarabinofuranosyl-QS-18 family components are present in relatively low amounts in extracts, meaning that they have not been subjected to detailed characterisation. Desarabinofuranosyl-QS-18 family components can be challenging to isolate from QS-21 family components. Desarabinofuranosyl-QS-18 family components include:desarabinofuranosyl-QS-18 2018 A component (i.e. triterpenoid glycosides identified as part of the ‘2018 Peak’ in FIG. 6). Suitably the desarabinofuranosyl-QS-18 2018 A component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.5 min, the primary component of the peak having a m / z of 2018 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-18 2018 A component may also be identified in the UPLC-UV methods described herein with a retention time of approximately 5.8 min. The desarabinofuranosyl-QS-18 2018 A component is believed to be identified in Kite 2004 as Peak 90 and corresponds to A-isomers of the xylose chemotype. Putative structures have been identified for the primary desarabinofuranosyl-QS-18 2018 A components using MS / MS. The desarabinofuranosyl-QS-18 2018 A component may consist of desarabinofuranosyl-QS-18 2018 A V1 (i.e. apiose isomer): and / or desarabinofuranosyl-QS-18 2018 A V2 (i.e. xylose isomer):desarabinofuranosyl-QS-18 1886 A component (i.e. triterpenoid glycosides identified as part of the ‘2018 Peak’ in FIG. 6). Suitably the desarabinofuranosyl-QS-18 1886 A component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.5 min and a m / z of 1886 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-18 1886 A component and corresponds to the A-isomer xylose chemotype may also be identified in the UPLC-UV methods described herein with a retention time of approximately 5.8 min. The desarabinofuranosyl-QS-18 1886 A component may consist of:desarabinofuranosyl-QS-18 2032 A component (i.e. triterpenoid glycosides identified as part of the ‘2018 Peak’ in FIG. 6). Suitably the desarabinofuranosyl-QS-18 2032 A component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.5 min and a m / z of 2032 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-18 2032 A component corresponds to A-isomers of the rhamnose chemotype and may also be identified in the UPLC-UV methods described herein with a retention time of approximately 5.8 min. The desarabinofuranosyl-QS-18 2032 A component may consist of desarabinofuranosyl-QS-18 2032 A V1 (i.e. apiose isomer): and / or desarabinofuranosyl-QS-18 2032 A V2 (i.e. xylose isomer):desarabinofuranosyl-QS-18 2018 B component (i.e. triterpenoid glycosides having a m / z of 2018 with negative mode electrospray mass spectrometry). The desarabinofuranosyl-QS-18 2018 B component corresponds to B-isomers of the xylose chemotype. Desarabinofuranosyl-QS-18 2018 B component may consist of desarabinofuranosyl-QS-18 2018 B V1 (i.e. apiose isomer): and / or desarabinofuranosyl-QS-18 2018 B V2 (i.e. xylose isomer):desarabinofuranosyl-QS-18 1886 B component (i.e. triterpenoid glycosides identified having a m / z of 1886 with negative mode electrospray mass spectrometry). The desarabinofuranosyl-QS-18 1886 B component corresponds to the B-isomer xylose chemotype. The desarabinofuranosyl-QS-18 1886 B component may consist of:desarabinofuranosyl-QS-18 2032 B component (i.e. triterpenoid glycosides having a m / z of 2032 with negative mode electrospray mass spectrometry). The desarabinofuranosyl-QS-18 2032 B component corresponds to B-isomers of the rhamnose chemotype. The desarabinofuranosyl-QS-18 2032 B component may consist of desarabinofuranosyl-QS-18 2032 B V1 (i.e. apiose isomer):and / or desarabinofuranosyl-QS-18 2032 B V (i.e. xylose isomer):acetylated desglucosyl-QS-17 family components (i.e. triterpenoid glycosides having alpha-O-rhamnosylation at the C2 position of the arabinofuranose moiety and acetylation of the C3 position of the fucose of QS-21 family components), such as:‘acetylated desglucosyl-QS-17 2176 A component’, being triterpenoid glycosides having a m / z of 2176 with negative mode electrospray mass spectrometry. The acetylated desglucosyl-QS-17 2176 A component corresponds to A-isomers of the xylose chemotype. The acetylated desglucosyl-QS-17 2176 A component may consist of acetylated desglucosyl-QS-17 2176 A V1 (i.e. apiose isomer): and / or acetylated desglucosyl-QS-17 2176 A V2 (i.e. xylose isomer):‘acetylated desglucosyl-QS-17 2044 A component’, being triterpenoid glycosides having m / z of 2044 with negative mode electrospray mass spectrometry. The acetylated desglucosyl-QS-17 2044 A component corresponds to the A-isomer xylose chemotype. The acetylated desglucosyl-QS-17 2044 A component may consist of acetylated desglucosyl-QS-17 2044 A:‘acetylated desglucosyl-QS-17 2190 A component’, being triterpenoid glycosides having a m / z of 2190 with negative mode electrospray mass spectrometry. The acetylated desglucosyl-QS-17 2190 A corresponds to the A-isomer rhamnose chemotype. The acetylated desglucosyl-QS-17 2190 A component may consist of acetylated desglucosyl-QS-17 2190 A V1 (i.e. apiose isomer): and / or acetylated desglucosyl-QS-17 2190 A V2 (i.e. xylose isomer):Starting saponins of direct relevance to the engineered glucosidase polypeptides for use in the methods of the invention are those having cleavable glucose residues, nevertheless, the engineered glucosidase polypeptides may be utilised in conjunction with additional enzymes capable of cleaving other sugar residues. Particular starting saponins of relevance to the engineered glucosidase polypeptides include:QS-18 family components;QS-17 family components; anddesarabinofuranosyl-QS-18 family components.Starting saponins of direct relevance to the engineered rhamnosidase polypeptides for use in the methods of the invention are those having cleavable rhamnose residues, nevertheless, the engineered rhamnosidase polypeptides may be utilised in conjunction with additional enzymes capable to cleaving other sugar residues. Particular starting saponins of relevance to the engineered rhamnosidase polypeptides include:desglucosyl-QS-17 family components;QS-17 family components; andacetylated desglucosyl-QS-17 family components.The methods of the present invention enzymatically modify a starting saponin obtained from a plant cell culture to provide a product saponin (i.e. a saponin resulting from an enzymatic modification).The product saponin may be a naturally occurring saponin (i.e. a steroid or terpenoid glycoside found in nature, though the product saponin is itself obtained by the methods of the invention) or an artificially created saponin (i.e. a steroid or terpenoid glycoside not found in nature).In some embodiments, the product saponin is a steroid glycoside. In other embodiments, the product saponin is a terpenoid glycoside, especially a triterpenoid glycoside.Naturally occurring product saponins may be saponins synthesized by plant cells cultured in vitro that originate from plants that produce saponins, such as the plants described below.Naturally occurring product saponins include those obtainable from, such as obtained from, an in vitro plant cell culture originating from the genera Gypsophilia, Saponaria, such as the Saponaria vaccaria species, or Saponaria officinalis species, or Quillaja (Bomford, 1992). Especially of interest are product saponins obtainable from an in vitro plant cell culture originating from Quillaja species. Particular product saponins of interest include those obtained from an in vitro plant cell culture originating from Quillaja brasiliensis or Quillaja saponaria. In one embodiment, the product saponin is obtained from an in vitro plant cell culture originating from Quillaja saponaria. In one embodiment, the product saponin is obtained from an in vitro plant cell culture originating from Quillaja brasiliensis. In certain embodiments, the product saponin is a quillaic acid glycoside.Product saponins obtainable from a culture of plant cells originating from Quillaja saponaria include:QS-18 family components (i.e. triterpenoid glycosides having beta-O-glucopyranosylation at the C3 position of the L-rhamnose moiety of QS-21 family components), such as:‘QS-18 2150 A component’. The QS-18 2150 A component may consist of QS-18 2150 A V1 (i.e. apiose isomer): and / or QS-18 2150 A V2 (i.e. xylose isomer):‘QS-18 2018 A component’. The QS-18 2018 A component may consist of QS-18 2018 A:‘QS-18 2164 A component’. The QS-18 2164 A component may consist of QS-18 2164 A V1 (i.e. apiose isomer): and / or QS-18 2164 A V2 (i.e. xylose isomer):‘QS-18 2150 B component’. The QS-18 2150 B component may consist of QS-18 2150 B V1 (i.e. apiose isomer): and / or QS-18 2150 B V2 (i.e. xylose isomer):‘QS-18 2018 B component’. The QS-18 2018 B component may consist of QS-18 2018 B:‘QS-18 2164 B component’. The QS-18 2164 B component may consist of QS-18 2164 B V1 (i.e. apiose isomer): and / or QS-18 2164 B V2 (i.e. xylose isomer):desglucosyl-QS-17 family components (i.e. triterpenoid glycosides having alpha-O-rhamnosylation at the C2 position of the arabinofuranose moiety of QS-21 family components), such as:‘desglucosyl-QS-17 2134 A component’. The desglucosyl-QS-17 2134 A component may consist of desglucosyl-QS-17 2134 A V1 (i.e. apiose isomer): and / or desglucosyl-QS-17 2134 A V2 (i.e. xylose isomer):‘desglucosyl-QS-17 2002 A component’. The desglucosyl-QS-17 2002 A component may consist of desglucosyl-QS-17 2002 A:‘desglucosyl-QS-17 2148 A component’. The desglucosyl-QS-17 2148 A component may consist of desglucosyl-QS-17 2148 A V1 (i.e. apiose isomer): and / or desglucosyl-QS-17 2148 A V2 (i.e. xylose isomer):‘desglucosyl-QS-17 2134 B component’. The desglucosyl-QS-17 2134 B component may consist of desglucosyl-QS-17 2134 B V1 (i.e. apiose isomer): and / or desglucosyl-QS-17 2134 B V2 (i.e. xylose isomer):‘desglucosyl-QS-17 2002 B component’. The desglucosyl-QS-17 2002 B component may consist of desglucosyl-QS-17 2002 B:‘desglucosyl-QS-17 2148 B component’. The desglucosyl-QS-17 2148 B component may consist of desglucosyl-QS-17 2148 B V1 (i.e. apiose isomer): and / or desglucosyl-QS-17 2148 B V2 (i.e. xylose isomer):QS-21 family components, such as:‘QS-21 1988 A component’, being the triterpenoid glycosides identified as part of the QS-21 main peak in FIG. 6 and having a m / z of 1988 with negative mode electrospray mass spectrometry. Suitably the QS-21 1988 A component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.4 min and a m / z of 1988 with negative mode electrospray mass spectrometry. The QS-21 1988 A component is believed to be identified in Kite 2004 as Peak 88 and corresponds to the A-isomer xylose chemotype structures S6 (apiose isomer) and S4 (xylose isomer) characterised in Nyberg 2000 and Nyberg 2003. The QS-21 1988 A component may consist of QS-21 1988 A V1 (i.e. apiose isomer): and QS-21 1988 A V2 (i.e. xylose isomer):‘QS-21 1856 A component’, being the triterpenoid glycosides identified as part of the QS-21 main peak in FIG. 6 and having a m / z of 1856 with negative mode electrospray mass spectrometry. Suitably the QS-21 1856 A component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.4 min and a m / z of 1856 with negative mode electrospray mass spectrometry. The QS-21 1856 A component is believed to be identified in Kite 2004 as Peak 86 and corresponds to the A-isomer xylose chemotype structure S2 characterised in Nyberg 2000 and Nyberg 2003. The QS-21 1856 A component may consist of:‘QS-21 2002 A component’, being the triterpenoid glycosides identified as part of the QS-21 main peak in FIG. 6 and having a m / z of 2002 with negative mode electrospray mass spectrometry. Suitably the QS-21 2002 A component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.4 min and a m / z of 2002 with negative mode electrospray mass spectrometry. The QS-21 2002 A component is believed to be identified in Kite 2004 as Peak 85 and corresponds to the A-isomer rhamnose chemotype of structures S3 and S5 characterised in Nyberg 2000 and Nyberg 2003. The QS-21 2002 A component may consist of QS-21 2002 A V1 (i.e. apiose isomer): and QS-21 2002 A V2 (i.e. xylose isomer):‘QS-21 1988 B component’, being the triterpenoid glycosides identified as part of the B-isomer peak in FIG. 6 and having a m / z of 1988 with negative mode electrospray mass spectrometry. Suitably the QS-21 1988 B component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.0 min and a m / z of 1988 with negative mode electrospray mass spectrometry. The QS-21 1988 B component corresponds to the B-isomer xylose chemotype structures S6a (apiose isomer) and S4a (xylose isomer) characterised in Nyberg 2000 and Nyberg 2003. The QS-21 1988 B component may consist of QS-21 1988 B V1 (i.e. apiose isomer): and QS-21 1988 B V2 (i.e. xylose isomer):‘QS-21 1856 B component’, being the triterpenoid glycosides identified as part of the B-isomer peak in FIG. 6 and having a m / z of 1856 with negative mode electrospray mass spectrometry. The QS-21 1856 B component corresponds to the B-isomer xylose chemotype structure S2a characterised in Nyberg 2000 and Nyberg 2003. The QS-21 1856 B component may consist of:‘QS-21 2002 B component’, being the triterpenoid glycosides having a m / z of 2002 with negative mode electrospray mass spectrometry. The QS-21 2002 B component corresponds to the B-isomer rhamnose chemotype of structures S3a and S5a characterised in Nyberg 2000 and Nyberg 2003. The QS-21 2002 B component may consist of QS-21 2002 B component V1 (i.e. apiose isomer): and QS-21 2002 B V2 (i.e. xylose isomer):desarabinofuranosyl-QS-21 family components (i.e. triterpenoid glycosides lacking the arabinofuranose moiety of QS-21 family components). The desarabinofuranosyl-QS-21 family components are present in relatively low amounts in extracts, meaning that they have not been subjected to detailed characterisation. Desarabinofuranosyl-QS-21 family components include:desarabinofuranosyl-QS-21 1856 A component (i.e. triterpenoid glycosides identified as part of the ‘Lyophilization Peak’ in FIG. 6). Suitably the desarabinofuranosyl-QS-21 1856 A component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.7 min, the primary component of the peak having a m / z of 1856 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-21 1856 A component is believed to be identified in Kite 2004 as Peak 96. Putative structures have been identified for the primary desarabinofuranosyl-QS-21 1856 A components using MS / MS. The desarabinofuranosyl-QS-21 1856 A component may consist of desarabinofuranosyl-QS-21 1856 A V1 (i.e. apiose isomer): and / or desarabinofuranosyl-QS-21 1856 A V2 (i.e. xylose isomer):desarabinofuranosyl-QS-21 1712 A component (i.e. triterpenoid glycosides identified as part of the ‘Lyophilization Peak’ in FIG. 6). Suitably the desarabinofuranosyl-QS-21 1712 A component in the UPLC-UV / MS methods described herein has a retention time of approximately 4.7 min and a m / z of 1712 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-21 1712 A component may consist of desarabinofuranosyl-QS-21 1712 A:desarabinofuranosyl-QS-21 1870 A component, i.e. triterpenoid glycosides having a m / z of 1870 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-21 1870 A component may consist of desarabinofuranosyl-QS-21 1870 A V1 (i.e. apiose isomer): and / or desarabinofuranosyl-QS-21 1870 A V2 (i.e. xylose isomer):desarabinofuranosyl-QS-21 1856 B component, i.e. triterpenoid glycosides having a m / z of 1856 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-21 1856 B component may consist of desarabinofuranosyl-QS-21 1856 B V1 (i.e. apiose isomer): and / or desarabinofuranosyl-QS-21 1856 B V2 (i.e. xylose isomer):desarabinofuranosyl-QS-21 1712 B component, i.e. triterpenoid glycosides having a m / z of 1712 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-21 1712 B component may consist of desarabinofuranosyl-QS-21 1712 B:desarabinofuranosyl-QS-21 1870 B component, i.e. triterpenoid glycosides having a m / z of 1870 with negative mode electrospray mass spectrometry. The desarabinofuranosyl-QS-21 1870 B component may consist of desarabinofuranosyl-QS-21 1870 B V1 (i.e. apiose isomer): and / or desarabinofuranosyl-QS-21 1870 B V2 (i.e. xylose isomer):acetylated QS-21 family components (i.e. triterpenoid glycosides having acetylation of the C3 position of the fucose of QS-21 family components), such as:‘acetylated QS-21 2030 A component’, being triterpenoid glycosides having a m / z of 2030 with negative mode electrospray mass spectrometry. The acetylated QS-21 2030 A corresponds to the A-isomer xylose chemotype. The acetylated QS-21 2030 A component may consist of acetylated QS-21 2030 A V1 (i.e. apiose isomer): and / or acetylated QS-21 2030 A V2 (i.e. xylose isomer):‘acetylated QS-21 1898 A component’, being triterpenoid glycosides having a m / z of 1898 with negative mode electrospray mass spectrometry. The acetylated QS-21 1898 A corresponds to the A-isomer xylose chemotype. The acetylated QS-21 1898 A component may consist of acetylated QS-21 1898 A:‘acetylated QS-21 2044 A component’, being triterpenoid glycosides having a m / z of 2044 with negative mode electrospray mass spectrometry. The acetylated QS-21 2044 A corresponds to A-isomers of the rhamnose chemotype. The acetylated QS-21 2044 A component may consist of acetylated QS-21 2044 A V1 (i.e. apiose isomer): and / or acetylated QS-21 2044 A V2 (i.e. xylose isomer):Product saponins of direct relevance to the engineered glucosidase polypeptides for use in the methods of the invention are those where a glucose residue has been cleaved relative to a starting saponin. Nevertheless, the engineered glucosidase polypeptides may be utilised in conjunction with additional enzymes capable to cleaving other sugar residues. Particular product saponins of relevance to the engineered glucosidase polypeptides include:desglucosyl-QS-17 family components;QS-21 family components; anddesarabinofuranosyl-QS-21 family components.Product saponins of direct relevance to the engineered rhamnosidase polypeptides for use in the methods of the invention are those where a rhamnose residue has been cleaved relative to a starting saponin. Nevertheless, the engineered rhamnosidase polypeptides may be utilised in conjunction with additional enzymes capable to cleaving other sugar residues. Particular product saponins of relevance to the engineered rhamnosidase polypeptides include:QS-18 family components;QS-21 family components; andacetylated QS-21 family components.The term ‘QS-18 family components’ as used herein means the xylose chemotype QS-18 2150 component (A and B isomers, and apiose and xylose isomers: QS-18 2150 A V1, QS-18 2150 A V2, QS-18 2150 B V1 and QS-18 2150 B V2), the xylose chemotype QS-18 2018 component (A and B isomers: QS-18 2018 A and QS-18 2018 B), the rhamnose chemotype QS-18 2164 component (A and B isomers, and apiose and xylose isomers: QS-18 2164 A V1, QS-18 2164 A V2, QS-18 2164 B V1 and QS-18 2164 B V2).The term ‘desglucosyl-QS-17 family components’ as used herein means the xylose chemotype desglucosyl-QS-17 2134 component (A and B isomers, and apiose and xylose isomers: desglucosyl-QS-17 2134 A V1, desglucosyl-QS-17 2134 A V2, desglucosyl-QS-17 2134 B V1 and desglucosyl-QS-17 2134 B V2), the xylose chemotype desglucosyl-QS-17 2002 component (A and B isomers: desglucosyl-QS-17 2002 A and desglucosyl-QS-17 2002 B), the rhamnose chemotype desglucosyl-QS-17 2148 component (A and B isomers, and apiose and xylose isomers: desglucosyl-QS-17 2148 A V1, desglucosyl-QS-17 2148 A V2, desglucosyl-QS-17 2148 B V1 and desglucosyl-QS-17 2148 B V2).The term ‘QS-17 family components’ as used herein means the xylose chemotype QS-17 2296 component (A and B isomers, and apiose and xylose isomers: QS-17 2296 A V1, QS-17 2296 A V2, QS-17 2296 B V1 and QS-17 2296 B V2), the xylose chemotype QS-17 2164 component (A and B isomers: QS-17 2164 A and QS-17 2164 B), the rhamnose chemotype QS-17 2310 component (A and B isomers, and apiose and xylose isomers: QS-17 2310 A V1, QS-172310 A V2, QS-17 2310 B V1 and QS-17 2310 B V2).The term ‘QS-21 family components’ as used herein means the xylose chemotype QS-21 1988 component (A and B isomers, and apiose and xylose isomers: QS-21 1988 A V1, QS-21 1988 A V2, QS-21 1988 B V1 and QS-21 1988 B V2), the xylose chemotype QS-21 1856 component (A and B isomers: QS-21 1856 A and QS-21 1856 B), the rhamnose chemotype QS-21 2002 component (A and B isomers, and apiose and xylose isomers: QS-21 2002 A V1, QS-21 2002 A V2, QS-21 2002 B V1 and QS-21 2002 B V2).The term ‘desarabinofuranosyl-QS-18 family components’ as used herein means the xylose chemotype desarabinofuranosyl-QS-18 2018 component (A and B isomers, and apiose and xylose isomers: desarabinofuranosyl-QS-18 2018 A V1, desarabinofuranosyl-QS-18 2018 A V2, desarabinofuranosyl-QS-18 2018 B V1 and desarabinofuranosyl-QS-18 2018 B V2), the xylose chemotype desarabinofuranosyl-QS-18 1886 component (A and B isomers: desarabinofuranosyl-QS-18 1886 A and desarabinofuranosyl-QS-18 1886 B), the rhamnose chemotype desarabinofuranosyl-QS-18 2032 component (A and B isomers, and apiose and xylose isomers: desarabinofuranosyl-QS-18 2032 A V1, desarabinofuranosyl-QS-18 2032 A V2, desarabinofuranosyl-QS-18 2032 B V1 and desarabinofuranosyl-QS-18 2032 B V2).The term ‘acetylated desglucosyl-QS-17 family components’ as used herein means xylose chemotype acetylated desglucosyl-QS-17 2176 component (apiose and xylose isomers: acetylated desglucosyl-QS-17 2176 A V1 and acetylated desglucosyl-QS-17 2176 A V2), the xylose chemotype acetylated desglucosyl-QS-17 2044 A component, the rhamnose chemotype acetylated desglucosyl-QS-17 2190 component (apiose and xylose isomers: acetylated desglucosyl-QS-17 2190 A V1 and acetylated desglucosyl-QS-17 2190 A V2).The term ‘desarabinofuranosyl-QS-21 family components’ as used herein means xylose chemotype desarabinofuranosyl-QS-21 1856 component (A and B isomers, and apiose and xylose isomers: desarabinofuranosyl-QS-21 1856 A V1, desarabinofuranosyl-QS-21 1856 A V2, desarabinofuranosyl-QS-21 1856 B V1 and desarabinofuranosyl-QS-21 1856 B V2), the xylose chemotype desarabinofuranosyl-QS-21 1712 component (A and B isomers: desarabinofuranosyl-QS-21 1712 A and desarabinofuranosyl-QS-21 1712 B), the rhamnose chemotype desarabinofuranosyl-QS-21 1870 component (A and B isomers, and apiose and xylose isomers: desarabinofuranosyl-QS-21 1870 A V1, desarabinofuranosyl-QS-21 1870 A V2, desarabinofuranosyl-QS-21 1870 B V1 and desarabinofuranosyl-QS-21 1870 B V2).The term ‘acetylated QS-21 family components’ as used herein means xylose chemotype acetylated QS-21 2030 component (apiose and xylose isomers: acetylated QS-21 2030 A V1 and acetylated QS-21 2030 A V2), the xylose chemotype acetylated QS-21 1898 A component, the rhamnose chemotype acetylated QS-21 2044 component (apiose and xylose isomers: acetylated QS-21 2044 A V1 and acetylated QS-21 2044 A V2).Plant Cell CultureThe term ‘plant cell culture’ is to be understood as the in vitro culture of any plant tissues or any plant cells derived from any tissue from plants producing saponins, such as e.g. plants described and referred to earlier. Tissues or cells from the plant cell culture are capable of synthesizing saponins. The term ‘capable of synthesizing saponins’, in the sense of the present invention, refers to the ability of cells to synthesize and produce saponins. Depending on different factors, such as e.g. the origin of the plant cell culture, the cell culture conditions, saponin synthesis and production thereof may require to be triggered. The term ‘under conditions leading to the synthesis of saponins’, in the sense of the present invention, refers to conditions triggering and resulting into the effective synthesis of saponins and production thereof.A plant cell culture in the sense of the present invention may be obtained and generated according to any known method. Lambert et al. (2011) provides a review on in vitro cultures of saponin-producing plants in general. WO 94 / 10291 (incorporated herein by reference) discloses how to obtain an in vitro culture of plant cells originating from Quillaja saponaria species that are capable of synthesizing and producing saponins.Methods for culturing plant cells, and general culture conditions for plant cells are known in the art. Conventional culture media known for plant cell culture, such as e.g. classical Murashige and Skoog (MS) medium, White's medium, or Linsmaler & Skoog's medium can be used in the methods of the invention. These media typically contain at least one or more macronutrients, e.g. selected from NH4NO3, KNO3, CaCl2), MgSO4, KH2PO4, NH4Cl, or KCl; at least one or more micronutrients, e.g. selected from KI, H3B03, MnSO4, ZnSO4, Na2MoO4, CuSO4, CoCl2, DeSO4, or Na2EDTA; at least one or more vitamins, e.g. selected from myoinisotol, nicotinic acid, pyrodixine-HCl, or thiamine-HCl, for example at a total concentration between 0.01 and 3 g / L, such as between 50 and 150 mg / L; at least one or more amino acids, such as glycine; at least one or more carbon source, e.g. selected from sucrose, glucose or fructose; and at least one or more plant hormone, e.g. selected from one or more cytokinins, or one or more auxins, such as 2,4-D and / or NAA.Plant cells used in the methods of the invention may originate from different tissues or organs of a given plant body, e.g. primordia, leaves, stems, hairy roots, internodes, cambium, whether cultured in suspension in a liquid medium or on a solid medium, e.g. calli. In one embodiment, the plant cells used in the method of the invention originate from the cambium, e.g. are cambial meristematic cells (CMC). In an alternative embodiment, the plant cells originate from hairy roots.Plant cells used in the methods of the invention may be a callus, e.g. deriving from the cambium of the plant. A “callus” is typically defined as a cluster of dedifferentiated cells cultured on solidified medium. Callus generation may be achieved from any plant tissue explant by any method known to the skilled person, e.g. the methods described in WO 94 / 10291, in US 2019 / 0134128 or in WO 15 / 082978. In brief, tissue explants from a plant of a small size may be surface sterilized, e.g. by washing thoroughly with clean water, using a disinfectant such as hypochlorite, using wet agents, such as Tween or Triton, using antibiotics and / or using anti-fungal agents. Surface sterilized explants are then, typically, laid on the surface of solidified medium, such as agar, and incubated in a sterile environment, until a mass of undifferentiated cells grows (typically between 2 to 12 weeks, e.g. 8 weeks) in proximity to the plant source material. Calli may be gradually purified and further propagated by means of repeating the similar solid medium-culturing, that is, by inoculating fresh solid medium by turns with small pieces of callus formed in the previous solid medium-culturing, e.g. every 4 weeks. Callus culture conditions including media components, such as MS media, pH ranges, carbon sources, nitrogen sources, macro-salts and micro-salts, vitamins and growth regulators are well known in the art.Calli thus formed and refined on the solid medium by subculture may be inoculated into a liquid medium and cultured so as to obtain a suspension cell culture. The terms “suspension plant culture” and “suspension of plant cells” are interchangeable and refer to an in vitro culture of plant cells dispersed in a liquid medium and grown in suspension. In one embodiment, the plant cells for use in the methods of the invention are grown in suspension in a liquid medium. To obtain plant cells into a suspension culture, cells are for example removed from a callus and transferred to sterile culture vessels containing nutrient culture medium. It is appreciated that optimized media for suspension cell lines may differ from the optimum for callus. It is within the ambit of the skilled person to determine suitable and optimal culture media.The transition from a callus to suspension cell cultures is also known to the skilled person (described e.g. in WO 94 / 10291 or US 2019 / 0134128). Suspension culture conditions including media components, pH ranges, carbon sources, nitrogen sources, macro-salts and micro-salts, vitamins and growth regulators are well known in the art, e.g. classical MS media. Once initiated and adapted to growth in suspension, suspension cells may be sub-cultured or propagated, for example by dilution, e.g. every 4 weeks.Saponin synthesis by the plant cells used in the methods of the invention (i.e. capable of synthesizing saponins) may be optimized by eliciting the cells with an elicitor. Suitable elicitors for use in the methods of the invention are monococarboxylic compound-type elicitors, such as 5-chlorosalicyclic acid, salicyclic acid, acetylsalicyclic acid, a methyl ester, e.g. methyl jasmonate (MeJa), or the chemically synthesized 2-HEJ. In one embodiment, the elicitor is MeJa.While the concentration of elicitors is typically referred to by reference to the volume of the culture medium, an alternative way to define the concentration is by reference to the PCV %. The term “PCV” stands for Packed Cell Volume and refers to the volume occupied by cells in culture medium. It may be calculated as follows: PCV (%)=(volume of cell pellet / volume of sample)×100. A suitable range of an elicitor, e.g. MeJa is from 0.5 to 10 μM / PCV %.Elicitors may be added directly to the culture medium. Alternatively, the culture medium may be replaced with a culture medium including the elicitor(s).Elicitation duration may be adjusted, depending on the plant cells under consideration. It is within the ambit of the skilled person to determine the optimal duration. Suitably, the elicitation may take place between 1 to 14 days, more suitably between 2 to 10 days, and even more suitably, between 3 to 8 days.Alternatively, or additionally, to elicitation, saponin synthesis by the plant cells used in the method of the invention may be further optimized by depleting the cells in nitrogen, e.g. prior to elicitation. Nitrogen depletion means reducing the level of any source of nitrogen present in the culture medium. Suitably, e.g. prior to elicitation, the cell culture medium of the cells is removed and replaced with a culture medium containing no source of nitrogen. Suitable ranges for the duration of nitrogen depletion (prior to elicitation) are from 1 to 9 days, suitably, from 2 to 7 days, and more suitably from 3 to 6 days.Elicitation and / or nitrogen depletion are non-limiting examples of ‘conditions leading to the synthesis of saponins’ in the sense of the present invention.Saponins synthesized by the plant cells used in the methods of the invention may either be extracellularly secreted into the culture medium and / or remain located intracellularly. Accordingly, a starting saponin intended to be enzymatically modified, or converted, in the sense of the present invention, may be included in the cell culture medium. In other words, the cell culture medium of plant cells capable of synthesizing saponins may be used as starting material, and subject to enzymatic treatment. In some embodiments, glycosidases are added to the culture medium, optionally after nitrogen has been depleted from the culture medium as described earlier and / or cells have been elicited with an elicitor as described earlier. Alternatively, a starting saponin intended to be enzymatically modified, or converted, in the sense of the present invention, may be recovered from the intracellular content of the plant cells. For example, the plant cells may be lysed (by any known in the art technique) and the resulting intracellular content be subject to enzymatic treatment.In the sense of the present invention, the term ‘plant cell culture extract’ is to be understood as any extract, or material, obtained from an in vitro plant cell culture, that includes saponins, whether synthesized saponins and / or recovered saponins, including the starting saponin and / or the product saponin. For example, a plant cell culture extract may be the plant cells, or the plant cells that have been lysed. A plant cell culture extract may be crude, or partially or wholly purified. A plant cell culture extract may be obtained from the genera Gypsophilia, Saponaria, such as Saponaria vaccaria or Saponaria officinalis or Quillaja (Bomford, 1992), such as a plant cell culture extract obtained from Quillaja species. Particular plant cell culture extracts include that obtained from Quillaja brasiliensis or Quillaja saponaria. In one embodiment, the plant cell culture extract is obtained from Quillaja saponaria. In one embodiment, the plant cell culture extract is obtained from Quillaja brasiliensis. The term ‘recover’ is to be understood as collecting saponins, for example saponins synthesized by the plant cells or the plant cell culture. Saponins may be recovered from the culture medium. Alternatively, saponins may be recovered from the plant cells, e.g. by extraction.Any known extraction method is suitable (e.g. as described in WO 94 / 10291), using non-aqueous polar solvents, extraction using an acid medium or a basic medium, or extraction by mechanically disrupting the plant cells, such as by ball milling, sonication. Alternatively, saponins may be extracted by freezing the cell pellet (resulting in cell lysis) obtained after centrifugation of the cell culture. Suitably, the cell pellet is frozen at −20° C., and more suitably at −70° C., e.g. at least for 24 hours.Extraction may be performed using water or lower alcohols (e.g. methanol or ethanol) as solvents, including mixtures thereof. In one embodiment, the starting saponin is obtained by aqueous extraction (e.g. using solvent comprising at least 80% v / v water, especially at least 90% v / v water, such as at least 95% v / v water). In one embodiment, the starting saponin is obtained by methanol extraction (e.g. using solvent comprising at least 80% v / v methanol, especially at least 90% v / v methanol, such as at least 95% v / v methanol). In one embodiment, the starting saponin is obtained by ethanol extraction (e.g. using solvent comprising at least 80% v / v ethanol, especially at least 90% v / v ethanol, such as at least 95% v / v ethanol). In one embodiment, the starting saponin is obtained by methanol / ethanol extraction (e.g. using solvent comprising at least 20% v / v methanol, especially at least 30% v / v methanol, such as at least 40% v / v methanol and at least 20% ethanol, especially at least 30% v / v ethanol, such as at least 40% v / v ethanol). In one embodiment, the starting saponin is obtained by water / ethanol extraction (e.g. using solvent comprising at least 20% v / v water, especially at least 30% v / v water, such as at least 40% v / v water and at least 20% ethanol, especially at least 30% v / v ethanol, such as at least 40% v / v ethanol. In one embodiment, the starting saponin is obtained by water / methanol extraction (e.g. using solvent comprising at least 20% v / v water, especially at least 30% v / v water, such as at least 40% v / v water and at least 20% methanol, especially at least 30% v / v methanol, such as at least 40% v / v methanol).Methods of the invention may be applied to starting saponin in a range of contexts. A starting saponin may be in the form of a minor component in a saponin-containing composition (ignoring solvents, if any), such as a minor component in a plant cell culture extract. A starting saponin may be in the form of a major component in a saponin containing composition, such as a major component in a plant cell culture extract. A starting saponin may be in the form of a minor component in a processed, such as partially purified, plant cell culture extract. A starting saponin may be in the form of a major component in a processed, such as partially purified, plant cell culture extract. In some embodiments, the starting saponin is substantially purified at the time of enzymatic modification. In further embodiments, the starting saponin is included in a crude material, such as a crude plant cell culture extract.Purification refers to the isolation of a component from other components. Partial purification therefore means the isolation of a component, to some degree, from other components. Substantial purification means the substantial isolation of a component from other components, such as wherein the component comprises at least 50% w / w, especially as at least 70%, particularly at least 80%, for example at least 90% of the component content (50%, 70%, 80% and 90% purity, respectively). Partial purification, in relation to a plant cell culture extract, means the isolation of the starting saponin, to some degree, from other extracted components. Substantially purified, in relation to a plant cell culture extract, means the substantial isolation of the starting saponin from other extracted components, such as wherein the starting saponin comprises at least 50% w / w, especially as at least 70%, particularly at least 80%, for example at least 90% of the extracted component content. Partial or substantial purification can be undertaken through various means including chromatography, filtration over semi-permeable membranes, treatment with selective adsorbants such as polyvinylpolypyrrolidone (PVPP) and the like.Although a starting saponin may be a specific chemical entity, in many circumstances involving saponins obtained by extraction, a plurality of starting saponins may be present, these being enzymatically modified to provide their corresponding product saponins. As mentioned above for individual saponins, the invention may be applied to a plurality of starting saponins in a range of contexts mutatis mutandis. A plurality of starting saponins comprising related starting saponins may undergo equivalent enzymatic modification concurrently. A plurality of starting saponins comprising distinguishable starting saponins may undergo different enzymatic modifications concurrently (in the presence of more than one enzyme) or in series (sequential treatment with separate enzymes). A plurality of starting saponins may contain both related and distinguishable starting saponins.Methods of the invention may be applied to a starting saponin in the form of a component of:crude plant cell culture extract, such as water and / or lower alcohol extract, especially aqueous cell extract;partially purified plant cell culture extract, such as water and / or lower alcohol plant cell culture extract, especially aqueous plant cell culture extract;PVPP treated plant cell culture extract, such as PVPP treated water and / or lower alcohol plant cell culture extract, especially PVPP treated aqueous plant cell culture extract;Plant cell culture fraction equivalent to Quil A;Plant cell culture fraction equivalent to Fraction A;Plant cell culture fraction equivalent to Fraction B (see Nyberg 2003);Plant cell culture fraction equivalent to Fraction C;QS-7 fraction;QS-17 fraction;QS-18 fraction; orQS-21 fraction.Methods of the invention may be applied to a starting saponin in a composition:QS-7 family components and QS-18 family components;QS-7 family components and QS-17 family components;QS-17 family components and QS-18 family components;QS-7 family components, QS-17 family components and QS-18 family components.As with other QS families, the QS-7 family components contain a plurality of related structures including xylose and rhamnose chemotypes, xylose and apiose isomers, A and B isomers:Certain QS-7 family components may lack glucose, or the rhamnose attached to the beta-D-fuc.Enzymatic ModificationsThe present invention provides the enzymatic modification of saponins obtained from an in vitro plant cell culture. Enzymatic modifications envisaged in the present invention include the conversion of a starting saponin into a product saponin by the removal of one or more sugar residues from the starting saponin. Suitably, the enzymatic modifications envisaged in the present invention are the conversion of a starting saponin into a product saponin by the removal of one or more sugar residues from the starting saponin.In certain embodiments, the enzymatic modification involves the removal of a single sugar residue i.e. removal of a terminal sugar residue (‘exo’ action) from a starting saponin. In other embodiments, enzymatic conversion involves the removal of a plurality of sugar residues from a starting saponin i.e. cleavage at a saccharide linkage other than in a terminal location (‘endo’ action), resulting in removal of a plurality of sugar residues (such as 2, 3 or 4 sugar residues) attached through said saccharide linkage.Particular sugar residues which may be removed comprise (such as consist of):glucose, in particular a terminal glucose, especially a beta-glucose, such as a beta-glucose from a quillaic acid glycoside, for example the beta-D-glucose residue highlighted below:rhamnose, in particular a terminal rhamnose, especially an alpha-rhamnose, such as an alpha-rhamnose from a quillaic acid glycoside, for example the alpha-L-rhamnose residues highlighted below:in particularParticular single sugar enzymatic conversions of interest include:QS-18 family components to QS-21 family components, such as:QS-18 2150 component (i.e. QS-18 2150 A and / or QS-18 2150 B) to QS-21 1988 component, such as:QS-18 2150 A component to QS-21 1988 A component, such as:QS-18 2150 A V1 to QS-21 1988 A V1QS-18 2150 A V2 to QS-21 1988 A V2QS-18 2150 B component to QS-21 1988 B component, such as:QS-18 2150 B V1 to QS-21 1988 B V1QS-18 2150 B V2 to QS-21 1998 B V2QS-18 2150 V1 component (i.e. QS-18 2150 A V1 and / or QS-18 2150 B V1) to QS-21 1988 V1 component, such as:QS-18 2150 A V1 to QS-21 1988 A V1QS-18 2150 B V1 to QS-21 1988 B V1QS-18 2150 V2 component (i.e. QS-18 2150 A V2 and / or QS-18 2150 B V2) to QS-21 1988 V2 component, such as:QS-18 2150 A V2 to QS-21 1988 A V2QS-18 2150 B V2 to QS-21 1988 B V2QS-18 2018 component (i.e. QS-18 2018 A and / or QS-18 2018 B) to QS-21 1856 component, such as:QS-18 2018 A component to QS-21 1856 A componentQS-18 2018 B component to QS-21 1856 B componentQS-18 2164 component (i.e. QS-18 2164 A and / or QS-18 2164 B) to QS-21 2002 component, such as:QS-18 2164 A component to QS-21 2002 A component, such as:QS-18 2164 A V1 to QS-21 2002 A V1QS-18 2164 A V2 to QS-21 2002 A V2QS-18 2164 B component to QS-21 2002 B component, such as:QS-18 2164 B V1 to QS-21 2002 B V1QS-18 2164 B V2 to QS-21 2002 B V2QS-18 2164 V1 component (i.e. QS-18 2164 A V1 and / or QS-18 2164 B V1) to QS-21 2002 V1 component, such as:QS-18 2164 A V1 to QS-21 2002 A V1QS-18 2164 B V1 to QS-21 2002 B V1QS-18 2164 V2 component (i.e. QS-18 2164 A V2 and / or QS-18 2164 B V2) to QS-21 2002 V2 component, such as:QS-18 2164 A V2 to QS-21 2002 A V2QS-18 2164 B V2 to QS-21 2002 B V2desglucosyl-QS-17 family components to QS-21 family components, such as:desglucosyl-QS-17 2134 component (i.e. desglucosyl-QS-17 2134 A and / or desglucosyl-QS-17 2134 B) to QS-21 1988 component, such as:desglucosyl-QS-17 2134 A component to QS-21 1988 A component, such as:desglucosyl-QS-17 2134 A V1 to QS-21 1988 A V1desglucosyl-QS-17 2134 A V2 to QS-21 1988 A V2desglucosyl-QS-17 2134 B component to QS-21 1988 B component, such as:desglucosyl-QS-17 2134 B V1 to QS-21 1988 B V1desglucosyl-QS-17 2134 B V2 to QS-21 1988 B V2desglucosyl-QS-17 2134 V1 component (i.e. desglucosyl-QS-17 2134 A V1 and / or desglucosyl-QS-17 2134 B V1) to QS-21 1988 V1 component, such as:desglucosyl-QS-17 2134 A V1 to QS-21 1988 A V1desglucosyl-QS-17 2134 B V1 to QS-21 1988 B V1desglucosyl-QS-17 2134 V2 component (i.e. desglucosyl-QS-17 2134 A V2 and / or desglucosyl-QS-17 2134 B V2) to QS-21 1988 V2 component, such as:desglucosyl-QS-17 2134 A V2 to QS-21 1988 A V2desglucosyl-QS-17 2134 B V2 to QS-21 1988 B V2desglucosyl-QS-17 2002 component (i.e. desglucosyl-QS-17 2002 A and / or desglucosyl-QS-17 2002 B) to QS-21 1856 component, such as:desglucosyl-QS-17 2002 A component to QS-21 1856 A componentdesglucosyl-QS-17 2002 B component to QS-21 1856 B componentdesglucosyl-QS-17 2148 component (i.e. desglucosyl-QS-17 2148 A and / or desglucosyl-QS-17 2148 B) to QS-21 2002 component, such as:desglucosyl-QS-17 2148 A component to QS-21 2002 A component, such as:desglucosyl-QS-17 2148 A V1 to QS-21 2002 A V1desglucosyl-QS-17 2148 A V2 to QS-21 2002 A V2desglucosyl-QS-17 2148 B component to QS-21 2002 B component, such as:desglucosyl-QS-17 2148 B V1 to QS-21 2002 B V1desglucosyl-QS-17 2148 B V2 to QS-21 2002 B V2desglucosyl-QS-17 2134 V1 component (i.e. desglucosyl-QS-17 2134 A V1 and / or desglucosyl-QS-17 2134 B V1) to QS-21 1988 V1 component, such as:desglucosyl-QS-17 2148 A V1 to QS-21 2002 A V1desglucosyl-QS-17 2148 B V1 to QS-21 2002 B V1desglucosyl-QS-17 2134 V2 component (i.e. desglucosyl-QS-17 2134 A V2 and / or desglucosyl-QS-17 2134 B V2) to QS-21 1988 V1 component, such as:desglucosyl-QS-17 2148 A V2 to QS-21 2002 A V2desglucosyl-QS-17 2148 B V2 to QS-21 2002 B V2QS-17 family components to QS-18 family components, such as:QS-17 2296 component (i.e. QS-17 2296 A and / or QS-17 2296 B) to QS-18 2150 component, such as:QS-17 2296 A component to QS-18 2150 A component, such as:QS-17 2296 A V1 to QS-18 2150 A V1QS-17 2296 A V2 to QS-18 2150 A V2QS-17 2296 B component to QS-18 2150 B component, such as:QS-17 2296 B V1 to QS-18 2150 B V1QS-17 2296 B V2 to QS-18 2150 B V2QS-17 2296 V1 component (i.e. QS-17 2296 A V1 and / or QS-17 2296 B V1) to QS-18 2150 V1 component, such as:QS-17 2296 A V1 to QS-18 2150 A V1QS-17 2296 B V1 to QS-18 2150 B V1QS-17 2296 V2 component (i.e. QS-17 2296 A V2 and / or QS-17 2296 B V1) to QS-18 2150 V2 component, such as:QS-17 2296 A V2 to QS-18 2150 A V2QS-17 2296 B V2 to QS-18 2150 B V2QS-17 2164 component (i.e. QS-17 2164 A and / or QS-17 2164 B) to QS-18 2018 component, such as:QS-17 2164 A component to QS-18 2018 A componentQS-17 2164 B component to QS-18 2018 B componentQS-17 2310 component (i.e. QS-17 2310 A and / or QS-17 2310 B) to QS-18 2164 component, such as:QS-17 2310 A component to QS-18 2164 A component, such as:QS-172310AV1 to QS-182164AV1QS-172310AV2 to QS-182164AV2QS-17 2310 B component to QS-18 2164 B component, such as:QS-17 2310 B V1 to QS-18 2164 B V1QS-17 2310 B V2 to QS-18 2164 B V2QS-17 2310 V1 component (i.e. QS-17 2310 A V1 and / or QS-17 2310 B V1) to QS-18 2164 V1 component, such as:QS-172310AV1 to QS-182164AV1QS-17 2310 B V1 to QS-18 2164 B V1QS-17 2310 V2 component (i.e. QS-17 2310 A V2 and / or QS-17 2310 B V2) to QS-18 2164 V2 component, such as:QS-172310AV2 to QS-182164AV2QS-17 2310 B V2 to QS-18 2164 B V2QS-17 family components to desglucosyl-QS-17 family components, such as:QS-17 2296 component (i.e. QS-17 2296 A and / or QS-17 2296 B) to desglucosyl-QS-17 2134 component, such as:QS-17 2296 A component to desglucosyl-QS-17 2134 A component, such as:QS-17 2296 A V1 to desglucosyl-QS-17 2134 A V1QS-17 2296 A V2 to desglucosyl-QS-17 2134 A V2QS-17 2296 B component to desglucosyl-QS-17 2134 B component, such as:QS-17 2296 B V1 to desglucosyl-QS-17 2134 B V1QS-17 2296 B V2 to desglucosyl-QS-17 2134 B V2QS-17 2296 V1 component (i.e. QS-17 2296 A V1 and / or QS-17 2296 B V1) to desglucosyl-QS-17 2134 V1 component, such as:QS-17 2296 A V1 to desglucosyl-QS-17 2134 A V1QS-17 2296 B V1 to desglucosyl-QS-17 2134 B V1QS-17 2296 V2 component (i.e. QS-17 2296 A V2 and / or QS-17 2296 B V1) to desglucosyl-QS-17 2134 V2 component, such as:QS-17 2296 A V2 to desglucosyl-QS-17 2134 A V2QS-17 2296 B V2 to desglucosyl-QS-17 2134 B V2QS-17 2164 component (i.e. QS-17 2164 A and / or QS-17 2164 B) to desglucosyl-QS-17 2002 component, such as:QS-17 2164 A component to desglucosyl-QS-17 2002 AQS-17 2164 B component to desglucosyl-QS-17 2002 BQS-17 2310 component (i.e. QS-17 2310 A and / or QS-17 2310 B) to desglucosyl-QS-17 2148 component, such as:QS-17 2310 A component to desglucosyl-QS-17 2148 A component, such as:QS-17 2310 A V1 to desglucosyl-QS-17 2148 A V1QS-17 2310 A V2 to desglucosyl-QS-17 2148 A V2QS-17 2310 B component to QS-21, such as:QS-17 2310 B V1 to desglucosyl-QS-17 2148 B V1QS-17 2310 B V2 to desglucosyl-QS-17 2148 B V2QS-17 2310 V1 component (i.e. QS-17 2310 A V1 and / or QS-17 2310 B V1) to desglucosyl-QS-17 2148 V1 component, such as:QS-17 2310 A V1 to desglucosyl-QS-17 2148 A V1QS-17 2310 B V1 to desglucosyl-QS-17 2148 B V1QS-17 2310 V2 component (i.e. QS-17 2310 A V2 and / or QS-17 2310 B V2) to desglucosyl-QS-17 2148 V2 component, such as:QS-17 2310 A V2 to desglucosyl-QS-17 2148 A V2QS-17 2310 B V2 to desglucosyl-QS-17 2148 B V2Other single sugar enzymatic conversions of interest include:desarabinofuranosyl-QS-18 family components to desarabinofuranosyl-QS-21 family components, such as:desarabinofuranosyl-QS-18 2018 component (i.e. desarabinofuranosyl-QS-18 2018 A and / or desarabinofuranosyl-QS-18 2018 B) to desarabinofuranosyl-QS-21 1856 component, such as:desarabinofuranosyl-QS-18 2018 A component to desarabinofuranosyl-QS-21 1856 A component, such as:desarabinofuranosyl-QS-18 2018 A V1 to desarabinofuranosyl-QS-21 1856 A V1desarabinofuranosyl-QS-18 2018 A V2 to desarabinofuranosyl-QS-21 1856 A V2desarabinofuranosyl-QS-18 2018 B component to desarabinofuranosyl-QS-21 1856 B component, such as:desarabinofuranosyl-QS-18 2018 B V1 to desarabinofuranosyl-QS-21 1856 B V1desarabinofuranosyl-QS-18 2018 B V2 to desarabinofuranosyl-QS-21 1856 B V2desarabinofuranosyl-QS-18 2018 V1 component (i.e. desarabinofuranosyl-QS-18 2018 A V1 and / or desarabinofuranosyl-QS-18 2018 B V1) to desarabinofuranosyl-QS-21 1856 V1 component, such as:

[1637] desarabinofuranosyl-QS-18 2018 A V1 to desarabinofuranosyl-QS-21 1856 A V1

[1638] desarabinofuranosyl-QS-18 2018 B V1 to desarabinofuranosyl-QS-21 1856 B V1

[1639] desarabinofuranosyl-QS-18 2018 V2 component (i.e. desarabinofuranosyl-QS-18 2018 A V2 and / or desarabinofuranosyl-QS-18 2018 B V2) to desarabinofuranosyl-QS-21 1856 V2 component, such as:

[1640] desarabinofuranosyl-QS-18 2018 A V2 to desarabinofuranosyl-QS-21 1856 A V2

[1641] desarabinofuranosyl-QS-18 2018 B V2 to desarabinofuranosyl-QS-21 1856 B V2

[1642] desarabinofuranosyl-QS-18 1886 component (i.e. desarabinofuranosyl-QS-18 1886 A and / or desarabinofuranosyl-QS-18 1886 B) to desarabinofuranosyl-QS-21 1712 component, such as:

[1643] desarabinofuranosyl-QS-18 1886 A component to desarabinofuranosyl-QS-21 1712 A

[1644] desarabinofuranosyl-QS-18 1886 B component to desarabinofuranosyl-QS-21 1712 B

[1645] desarabinofuranosyl-QS-18 2032 component (i.e. desarabinofuranosyl-QS-18 2032 A and / or desarabinofuranosyl-QS-18 2032 B) to desarabinofuranosyl-QS-21 1870 component, such as:

[1646] desarabinofuranosyl-QS-18 2032 A component to desarabinofuranosyl-QS-21 1870 A component, such as:

[1647] desarabinofuranosyl-QS-18 2032 A V1 to desarabinofuranosyl-QS-21 1870 A V1

[1648] desarabinofuranosyl-QS-18 2032 A V2 to desarabinofuranosyl-QS-21 1870 A V2

[1649] desarabinofuranosyl-QS-18 2032 B component to QS-21, such as:

[1650] desarabinofuranosyl-QS-18 2032 B V1 to desarabinofuranosyl-QS-21 1870 B V1

[1651] QS-17 desarabinofuranosyl-QS-18 2032 B V2 to desarabinofuranosyl-QS-21 1870 B V2

[1652] desarabinofuranosyl-QS-18 2032 V1 component (i.e. desarabinofuranosyl-QS-18 2032 A V1 and / or desarabinofuranosyl-QS-18 2032 B V1) to desarabinofuranosyl-QS-21 1870 V1 component, such as:

[1653] desarabinofuranosyl-QS-18 2032 A V1 to desarabinofuranosyl-QS-21 1870 A V1

[1654] desarabinofuranosyl-QS-18 2032 B V1 to desarabinofuranosyl-QS-21 1870 B V1

[1655] desarabinofuranosyl-QS-18 2032 V2 component (i.e. desarabinofuranosyl-QS-18 2032 A V2 and / or desarabinofuranosyl-QS-18 2032 B V2) to desarabinofuranosyl-QS-21 1870 V2 component, such as:

[1656] desarabinofuranosyl-QS-18 2032 A V2 to desarabinofuranosyl-QS-21 1870 A V2

[1657] desarabinofuranosyl-QS-18 2032 B V2 to desarabinofuranosyl-QS-21 1870 B V2

[1658] acetylated desglucosyl-QS-17 components to acetylated QS-21 family components, such as:

[1659] acetylated desglucosyl-QS-17 2176 A to acetylated QS-21 2030 A, such as:

[1660] acetylated desglucosyl-QS-17 2176 A V1 to acetylated QS-21 2030 A V1

[1661] acetylated desglucosyl-QS-17 2176 A V2 to acetylated QS-21 2030 A V2

[1662] acetylated desglucosyl-QS-17 2044 A to acetylated QS-21 1898 A component

[1663] acetylated desglucosyl-QS-17 2190 A to acetylated QS-21 2044 A, such as:

[1664] acetylated desglucosyl-QS-17 2190 A V1 to acetylated QS-21 2044 A V1

[1665] acetylated desglucosyl-QS-17 2190 A V2 to acetylated QS-21 2044 A V2.

[1666] Enzymatic conversions may be applied to a single starting saponin or a plurality of starting saponins in parallel. It will be appreciated that a process may comprise or consist of the conversions specified above, depending on the composition of the starting material and the enzymes used. Furthermore, while a process may be limited to the use of a single enzyme intended to remove a particular sugar residue or group of sugar residues from (i) a single starting saponin, (ii) a family of starting saponins, or (iii) from a plurality of families of starting saponins; processes may also use a plurality of enzymes intended to remove a plurality of sugar residues from (i) a single starting saponin, (ii) a family of starting saponins, or (iii) from a plurality of families of starting saponins. Processes involving multiple enzymes may be undertaken in series (i.e. a single enzyme is applied to saponin material at any time) or in parallel (i.e. more than one enzyme is applied to saponin material at any time, such as two or three enzymes, in particular two enzymes), or combinations thereof.

[1667] Processes involving the removal of multiple sugar residues may involve the removal of single (but different) sugar residues from multiple starting saponins and / or the removal of multiple sugar residues from particular starting saponins (such as 2, 3 or 4 residues, in particular 2 or 3, especially 2 residues). Removal of multiple sugar residues from particular starting saponins may involve any combination of removal of single residues and / or removal of a plurality of residues in a single cleavage.

[1668] Exemplary processes may comprise (such as consist of) the removal of glucose and rhamnose, in particular an alpha-rhamnose residue and a beta-glucose residue, such as the alpha-L-rhamnose residue and the beta-D-glucose residue from quillaic acid glycosides:

[1669] Particular multi-sugar enzymatic conversions of interest include:

[1670] QS-17 family components to QS-21 family components, such as:

[1671] QS-17 2296 component (i.e. QS-17 2296 A and / or QS-17 2296 B) to QS-21 1988 component, such as:

[1672] QS-17 2296 A component to QS-21 1988 A component, such as:

[1673] QS-17 2296 A V1 to QS-21 1988 A V1

[1674] QS-17 2296 A V2 to QS-21 1988 A V2

[1675] QS-17 2296 B component to QS-21 1988 B component, such as:

[1676] QS-17 2296 B V1 to QS-21 1988 B V1

[1677] QS-17 2296 B V2 to QS-21 1988 B V2

[1678] QS-17 2296 V1 component (i.e. QS-17 2296 A V1 and / or QS-17 2296 B V1) to QS-21 1988 V1 component, such as:

[1679] QS-17 2296 A V1 to QS-21 1988 A V1

[1680] QS-17 2296 B V1 to QS-21 1988 B V1

[1681] QS-17 2296 V2 component (i.e. QS-17 2296 A V2 and / or QS-17 2296 B V2) to QS-21 1988 V2 component, such as:

[1682] QS-17 2296 A V2 to QS-21 1988 A V2

[1683] QS-17 2296 B V2 to QS-21 1988 B V2

[1684] QS-17 2164 component (i.e. QS-17 2164 A and / or QS-17 2164 B) to QS-21 1856 component, such as:

[1685] QS-17 2164 A component to QS-21 1856 A component

[1686] QS-17 2164 B component to QS-21 1856 B component

[1687] QS-17 2310 component (i.e. QS-17 2310 A and / or QS-17 2310 B) to QS-21 2002 component, such as:

[1688] QS-17 2310 A component to QS-21 2002 A component, such as:

[1689] QS-17 2310 A V1 to QS-21 2002 A V1

[1690] QS-17 2310 A V2 to QS-21 2002 A V2

[1691] QS-17 2310 B component to QS-21 2002 B component, such as:

[1692] QS-17 2310 B V1 to QS-21 2002 B V1

[1693] QS-17 2310 B V2 to QS-21 2002 B V2

[1694] QS-17 2310 V1 component (i.e. QS-17 2310 A V1 and / or QS-17 2310 B V1) to QS-21 2002 V1, such as:

[1695] QS-17 2310 A V1 to QS-21 2002 A V1

[1696] QS-17 2310 B V1 to QS-21 2002 B V1

[1697] QS-17 2310 V2 component (i.e. QS-17 2310 A V2 and / or QS-17 2310 B V2) to QS-21 2002 V2, such as:

[1698] QS-17 2310 A V2 to QS-21 2002 A V2

[1699] QS-17 2310 B V2 to QS-21 2002 B V2.

[1700] Plant cell culture extracts may contain complex mixtures of saponin components and consequently may experience a plurality of conversions when multiple enzymes are present. For example, a starting mixture containing QS-17, QS-18 and desglucosyl-QS-17 components which is treated with an appropriate beta-glucosidase and alpha-rhamnosidase in parallel may undergo conversions including:

[1701] QS-18 family components to QS-21 family components, especially QS-18 2150 component to QS-21 1988 component;

[1702] desglucosyl-QS-17 family components to QS-21 family components, especially desglucosyl-QS-17 2134 component to QS-21 1988 component;

[1703] QS-17 family components to desglucosyl-QS-17 family components to QS-21 components, especially QS-17 2296 component to desglucosyl-QS-17 2134 component to QS-21 1988 component; and

[1704] QS-17 family components to QS-18 family components to QS-21 family components, especially QS-17 2296 component to QS-18 2150 component to QS-21 1988 component.Enzyme Selection

[1705] Extensive protein or DNA databases of natural and artificial glycosidases are available. Candidate enzymes may be selected and screened to assess suitability for achieving a particular conversion under particular reaction conditions. Suitability of an enzyme will depend on a number of factors including:

[1706] target sugar (e.g. glucose, rhamnose)

[1707] target sugar anomer (alpha or beta);

[1708] target sugar enantiomer (D or L);

[1709] target sugar location (endo or exo); and

[1710] target sugar environment (e.g. chemical / physical, impacting accessibility and reactivity).

[1711] Additional factors which facilitate effective conversions include:

[1712] rate of conversion;

[1713] environmental sensitivity—including pH, temperature, substrate, product and contaminant concentration tolerance; and

[1714] specificity for target sugar, including in respect of other sugar residues, other anomers, other sugar residue locations, and between different residues of the same sugar anomer and location within a substrate (if multiple such residues are present).

[1715] Those skilled in the art will appreciate that the level and type of specificity required of an enzyme will depend on the objective to be achieved and the general circumstances.

[1716] Conversion of QS-18 family components to QS-21 family components requires an enzyme demonstrating beta exo glucosidase activity.

[1717] Conversion of QS-17 family components to desglucosyl-QS-17 family components requires an enzyme demonstrating beta exo glucosidase activity.

[1718] Conversion of desglucosyl-QS-17 family components to QS-21 family components requires an enzyme demonstrating alpha exo rhamnosidase activity.

[1719] Conversion of QS-17 family components to QS-18 family components requires an enzyme demonstrating alpha exo rhamnosidase activity.

[1720] It may be noted that many Quillaja saponaria starting saponins of interest contain only one glucose residue. Many Quillaja saponaria starting saponins of interest contain a plurality of rhamnose residues, therefore selectivity for specific rhamnose residues is of greater importance practically. For example, conversion of desglucosyl-QS-17 family components to QS-21 components or QS-17 family components to QS-18 family components requires specificity for exo-rhamnosidase action over endo-rhamnosidase action. Furthermore, rhamnosidase specificity for the alpha-O-rhamnosylation at the C2 position of the arabinofuranose moiety over other terminal rhamnose residues (e.g. in the rhamnose chemotype components) may also be desirable. In certain embodiments it may be desirable to remove the terminal rhamnose from rhamnose chemotype components (alone or in conjunction with any alpha-O-rhamnosylation at the C2 position of the arabinofuranose moiety), to better facilitate their chromatographic separation from xylose chemotype components.

[1721] In one embodiment, enzymatic conversion is carried out by a single enzyme. The single enzyme may be a glucosidase, in particular a beta exo glucosidase. A single enzyme glucosidase may be an engineered glucosidase polypeptide described in the present invention. Alternatively, the single enzyme is a rhamnosidase, in particular an alpha exo rhamnosidase. A single enzyme rhamnosidase may be an engineered rhamnosidase polypeptide described in the present invention.

[1722] Preferred enzymes are those which efficiently enzymatically convert a starting saponin(s) to the desired product saponin(s) while demonstrating limited or no undesired conversion(s) of other saponin components present.

[1723] In one embodiment, enzymatic conversion to enzymatic is carried out by more than one enzyme, such as by two or three enzymes, especially by two enzymes. Enzymatic modification (or conversion) by more than one enzyme may involve sequential / series enzymatic modification. Alternatively, enzymatic modification (or conversion) by more than one enzyme may involve concurrent / parallel enzymatic modification. Enzymatic modification (or conversion) by at least three enzymes may involve a combination of sequential / series (modification by one enzyme) and concurrent / parallel (modification by at least two other enzymes) enzymatic modification, in any order. Where a plurality of enzymes are provided, these may be as distinct proteins or may be in the form of one or more fusion proteins.

[1724] An enzyme of interest is a glucosidase, such as a beta exo glucosidase. A glucosidase may be an engineered glucosidase polypeptide of the present invention. Another enzyme of interest is a rhamnosidase, such as an alpha exo rhamnosidase. A rhamnosidase may an engineered rhamnosidase polypeptide of the present invention. Enzyme combinations of interest include those comprising, such as consisting of, a glucosidase and a rhamnosidase, in particular a beta exo glucosidase and an alpha exo rhamnosidase. Enzymatic modification involving a glucosidase and a rhamnosidase, in particular a beta exo glucosidase and an alpha exo rhamnosidase, may be undertaken: sequentially with glucosidase (e.g. beta exo glucosidase) followed by rhamnosidase (e.g. alpha exo rhamnosidase), sequentially with rhamnosidase (e.g. alpha exo rhamnosidase) followed by glucosidase (e.g. beta exo glucosidase) or, conveniently, concurrently with both glucosidase (e.g. beta exo glucosidase) and rhamnosidase (e.g. alpha exo rhamnosidase). Particular enzyme combinations of interest are those comprising, such as consisting of, an engineered glucosidase of the present invention and an engineered rhamnosidase polypeptide of the present invention.

[1725] Enzymes utilised will typically be of external origin to saponin material i.e. not naturally found within the source of saponins obtained by extraction.

[1726] Enzymes may be native, i.e. naturally occurring glycosidases, or alternatively may be non-naturally occurring glycosidases. In one embodiment, a glucosidase enzyme is a naturally occurring glucosidase (e.g. exo glucosidase, such as beta exo glucosidase). In a second embodiment, a glucosidase enzyme is a non-naturally occurring glucosidase (e.g. exo glucosidase, such as beta exo glucosidase). In one embodiment, a rhamnosidase enzyme is a naturally occurring rhamnosidase (e.g. exo rhamnosidase, such as alpha exo rhamnosidase). In a second embodiment a rhamnosidase enzyme is a non-naturally occurring rhamnosidase (e.g. exo rhamnosidase, such as alpha exo rhamnosidase).

[1727] Enzymes may be modified relative to a reference enzyme (‘engineered’). Point mutations, either singly or in combination, introduced by engineering may provide benefits such as increased activity, increased specificity, increased stability, increased expression or other the like. Assays to confirm the properties of the enzymes are well known to those skilled in the field. For example, activity may be quantified by methods such as those shown in the examples (see Examples 4 to 7) or by analogous methods.

[1728] Different enzymes may show different sensitivity to environmental conditions, such as pH, temperature, substrate concentration, product concentration, solvent composition, presence of contaminants and the like. Such parameters may be taken into consideration during screening of candidate enzymes for the desired activity.

[1729] Candidate enzymes having beta glucosidase activity include those in EC3.2.1.21.

[1730] Beta exo glucosidases of interest include those described in Table 7, especially SEQ ID Nos. 262, 208, 63, 229, 250, 5, 101, 207, 169, 247, 302, 324, 319, 9, 240, 325 and 338, and functional variants thereof. Particular beta exo glucosidases of interest include SEQ ID Nos. 262, 208, 63, 229, 250, 5, 101, 207, 169, 247, 302, 324 and 319, and functional variants thereof, such as SEQ ID Nos. 262, 208, 63, 229, 250, 5, 101 and 207, and functional variants thereof.

[1731] Another group of beta exo glucosidases of interest include those described in Table 9, especially SEQ ID Nos. 850, 879, 868, 826, 804, 888, 881, 891, 816, 827, 857, 853, 842, 814, 886, 885, 838, 829, 808, 828, 870, 873, 844, 882, 874, 825, 824, 823, 810, 894, 849, 803, 890, 841, 832, 830, 845, 871, 837, 883 and 809, and functional variants thereof. Particular beta exo glucosidases of interest include SEQ ID Nos. 850, 879, 868, 826, 804, 888, 881, 891, 816, 827, 857, 853, 842, 814, 886, 885, 838, 829, 808, 828, 870, 873, 844, 882, 874, 825, 824, 823, 810, 894, 849, 803, 890 and 841, and functional variants thereof, such as SEQ ID Nos. 850, 879, 868, 826, 804, 888, 881, 891, 816, 827, 857, 853, 842, 814, 886, 885, 838, 829, 808, 828, 870, 873, 844, 882, 874, 825, 824, 823, 810 and 894, and functional variants thereof.

[1732] SEQ ID No. 262, and functional variants thereof, are particularly desirable beta exo glucosidases. In one embodiment the beta exo glucosidase comprises, such as consists of: (i) SEQ ID. 262; or (ii) a functional variant thereof having at least 80% identity to SEQ ID. 262, especially at least 90%, in particular at least 95%, such as at least 96%, at least 97%, at least 98%, for example at least 99% identity; or (iii) a functional fragment of at least 100, especially at least 200, particularly at least 300, such as at least 400, for example at least 500 contiguous amino acids of SEQ ID. 262.

[1733] Candidate enzymes having alpha rhamnosidase activity include those in EC3.2.1.40.

[1734] Alpha exo rhamnosidases of interest include SEQ ID Nos. 992, 1003, 1052, 1073, 1017, 1055, 1075, 1001, 1007, 1061, 1079, 1027, 1039, 1041, 989, 1053, 1018, 1066, 1082, 1076, 993, 1077, 1046, 1015, 1063, 1054, 1074, 1067 and 1033, and functional variants thereof. Particular alpha exo rhamnosidases of interest include SEQ ID Nos. 992, 1003, 1052, 1073, 1017, 1055, 1075, 1001, 1007, 1061, 1079, 1027, 1039, 1041, 989, 1053, 1018, 1066, 1082, 1076, 993 and 1077, and functional variants thereof, such as SEQ ID Nos. 992, 1003, 1052, 1073, 1017, 1055, 1075, 1001, 1007, 1061, 1079, 1027, 1039, 1041 and 989, and functional variants thereof.

[1735] SEQ ID No. 1017, and functional variants thereof, are particularly desirable exo rhamnosidases. In one embodiment, the alpha exo rhamnosidase comprises, such as consists of: (i) SEQ ID. 1017; or (ii) a functional variant thereof having at least 80% identity to SEQ ID. 1017, especially at least 90%, in particular at least 95%, such as at least 96%, at least 97%, at least 98%, for example at least 99% identity; or (iii) a functional fragment of at least 100, especially at least 200, particularly at least 300, such as at least 400, for example at least 500 contiguous amino acids of SEQ ID. 1017.

[1736] Functional variants of interest in the present application include those comprising, such as consisting of: (i) a sequence having at least 80% identity to the reference sequence, especially at least 90%, in particular at least 95%, such as at least 96%, at least 97%, at least 98%, for example at least 99% identity; or (ii) a fragment of at least 100, especially at least 200, particularly at least 300, such as at least 400, for example at least 500 contiguous amino acids of the reference sequence.

[1737] Certain desirable functional variants of interest include those comprising, such as consisting of, a sequence having 1 to 20 additions, deletions and / or substitutions relative to the reference sequence, especially 1 to 15 additions, deletions and / or substitutions, particularly 1 to 10 additions, deletions and / or substitutions, such as 1 to 5 additions, deletions and / or substitutions.

[1738] The degree of sequence identity may be determined using by the homology alignment algorithm of Needleman and Wunsch, the ClustalW program or the BLASTP algorithm, using default settings. An algorithm using global alignment (Needleman and Wunsch) is preferred.

[1739] “Percentage of sequence identity,”“percent identity,” and “percent identical” are used herein to refer to comparisons between polynucleotide sequences or polypeptide sequences, and are determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which either the identical nucleic acid base or amino acid residue occurs in both sequences or a nucleic acid base or amino acid residue is aligned with a gap to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity. Determination of optimal alignment and percent sequence identity is performed using the BLAST and BLAST 2.0 algorithms (see, e.g., Altschul, 1990; Altschul, 1997). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information website. Briefly, the BLAST analyses involve first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as, the neighborhood word score threshold (Altschul, supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, an expectation (E) of 10, M=5, N=−4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff, 1989).

[1740] Numerous other algorithms are available that function similarly to BLAST in providing percent identity for two sequences. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith, 1981, by the homology alignment algorithm of Needleman, 1970, by the search for similarity method of Pearson, 1988, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the GCG Wisconsin Software Package), or by visual inspection (see generally, Current Protocols in Molecular Biology, 1995)). Additionally, determination of sequence alignment and percent sequence identity can employ the BESTFIT or GAP programs in the GCG Wisconsin Software package (Accelrys, Madison WI), using default parameters provided. The ClustalW program is also suitable for determining identity.

[1741] Modestobacter marinus glucosidase (Uniparc reference UPI000260A2FA, Uniprot reference I4EYD5—SEQ ID No. 262 herein) is a naturally occurring glucosidase demonstrating beta exo glucosidase activity and, for example, is capable of the conversion of QS-18 family components to QS-21 family components. Despite its potent activity, the present inventors have found that the properties of wild type Modestobacter marinus glucosidase may be altered by the introduction of one or more mutations.

[1742] The present invention describes an engineered glucosidase polypeptide for use in the methods of the invention comprising, such as consisting of, an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID No. 262, or a functional fragment thereof, wherein the engineered glucosidase polypeptide includes at least one residue substitution from:

[1743] F44Y;

[1744] V60L;

[1745] G117A;

[1746] F170N;

[1747] V263G or V263L;

[1748] N351H or N351Q;

[1749] A355H, A355I, A355L, A355M, A355R, A355T or A355W;

[1750] A356P;

[1751] R357A, R357C, R357K, R357M or R357Q;

[1752] G362C;

[1753] T365A, T365N or T365S;

[1754] L367C;

[1755] V394R;

[1756] V395Y;

[1757] Q396E, Q396G, Q396N, Q396P, Q396R, Q396S or Q396Y;

[1758] F430W;

[1759] R435F;

[1760] V438T;

[1761] V440F;

[1762] F442M or F442Q;

[1763] G444T;

[1764] A473F or A473R;

[1765] L474C, L474I or L474V;

[1766] I475F;

[1767] L492C, L492G, L492H, L492I, L492N, L492Q, L492V, L492W or L492Y;

[1768] Q493F or Q493H;

[1769] P494H or P494I;

[1770] S495I, S495K or S495Q;

[1771] G496P or G496W;

[1772] D498A, D498E, D498F, D498I, D498K, D498L, D498N, D498P, D498R, D498S, D498T or D498V;

[1773] A502R;

[1774] M504G or M504R;

[1775] L507A or L507R;

[1776] T508M;

[1777] L529M;

[1778] F535P;

[1779] A536D or A536E;

[1780] A537R;

[1781] F541A, F541I, F541L, F541M or F541V;

[1782] L542I;

[1783] Q543G or Q543L;

[1784] E547L; and

[1785] Y585W.

[1786] The glucosidases will contain one to forty-two of the substitutions, such as two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, twenty-five, twenty-six to thirty or thirty-one to forty-three substitutions.

[1787] The present invention also describes an engineered glucosidase polypeptide for use in the methods of the invention comprising, such as consisting of, an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID No. 262, or a functional fragment thereof, wherein the engineered glucosidase polypeptide includes at least one residue substitution from:

[1788] F44Y;

[1789] V263L;

[1790] N351H;

[1791] A355H, A355M or A355W;

[1792] R357M;

[1793] T365N;

[1794] L367C;

[1795] Q396R;

[1796] V438T;

[1797] F442Q;

[1798] L474C;

[1799] I475F;

[1800] L492V, L492N or L492H,

[1801] M504R;

[1802] L507R; and

[1803] F541I.

[1804] The glucosidases will contain one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen or all sixteen substitutions.

[1805] The engineered glucosidase polypeptide may comprise, such as consist of, an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID No. 262, or a functional fragment thereof, wherein the engineered glucosidase polypeptide includes at least one residue substitution from: F44Y, V263L, A355W, R357M, T365N, L367C, Q396R, F442Q, L474C, I475F and F541I. Suitably the engineered glucosidase polypeptide comprises, such as consists of, an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID No. 262, or a functional fragment thereof, wherein the engineered glucosidase polypeptide includes the residue substitutions: F44Y, V263L, A355W, R357M, T365N, L367C, Q396R, F442Q, L474C, I475F and F541I.

[1806] The present invention describes a polypeptide for use in the methods of the invention comprising an amino acid sequence of sequence of SEQ ID No. 262 with one to twenty-five mutations selected from the list consisting of:

[1807] (i) F44Y

[1808] (ii) V263L

[1809] (iii) N351H

[1810] (iv) A355H, A355I, A355L, A355M, A355R, A355T or A355W

[1811] (v) A356P

[1812] (vi) R357M

[1813] (vii) T365N

[1814] (viii) L367C

[1815] (ix) F442Q

[1816] (x) G443D

[1817] (xi) A473F

[1818] (xii) L474C

[1819] (xiii) I475F

[1820] (xiv) L492H, L492N, L492V

[1821] (xv) P494I

[1822] (xvi) G496P

[1823] (xvii) D498P

[1824] (xviii) M504R

[1825] (xix) L507R

[1826] (xx) F535P

[1827] (xxi) A537R

[1828] (xxii) F541I

[1829] (xxiii) L542I

[1830] (xxiv) E547L and

[1831] (xxv) E588K.

[1832] Variant glucosidases will contain one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four or all twenty-five mutations.

[1833] In some embodiments, an engineered glucosidase is not a polypeptide comprising an amino acid sequence of sequence of SEQ ID No. 262 with one to twenty-five mutations selected from the list consisting of:

[1834] (i) F44Y

[1835] (ii) V263L

[1836] (iii) N351H

[1837] (iv) A355H, A355I, A355L, A355M, A355R, A355T or A355W

[1838] (v) A356P

[1839] (vi) R357M

[1840] (vii) T365N

[1841] (viii) L367C

[1842] (ix) F442Q

[1843] (x) G443D

[1844] (xi) A473F

[1845] (xii) L474C

[1846] (xiii) I475F

[1847] (xiv) L492H, L492N, L492V

[1848] (xv) P494I

[1849] (xvi) G496P

[1850] (xvii) D498P

[1851] (xviii) M504R

[1852] (xix) L507R

[1853] (xx) F535P

[1854] (xxi) A537R

[1855] (xxii) F541I

[1856] (xxiii) L542I

[1857] (xxiv) E547L and

[1858] (xxv) E588K.

[1859] The above-mentioned engineered glucosidase polypeptides may also be referred to herein as examples of ‘variant glucosidases’.

[1860] A variant glucosidase may contain F44Y.

[1861] A variant glucosidase may contain V60L.

[1862] A variant glucosidase may contain G117A.

[1863] A variant glucosidase may contain F170N.

[1864] A variant glucosidase may contain V263G or V263L, in particular V263L.

[1865] A variant glucosidase may contain N351H or N351Q, in particular N351H.

[1866] A variant glucosidase may contain A355H, A355I, A355L, A355M, A355R, A355T or A355W. In some embodiments, a variant glucosidase contains A355H. In some embodiments a variant glucosidase contains A355I. In some embodiments a variant glucosidase contains A355L. In some embodiments, a variant glucosidase contains A355M. In some embodiments a variant glucosidase contains A355R. In some embodiments a variant glucosidase contains A355T. In some embodiments, a variant glucosidase contains A355W.

[1867] A variant glucosidase may contain A356P.

[1868] A variant glucosidase may contain R357A, R357C, R357K, R357M or R357Q, in particular R357M.

[1869] A variant glucosidase may contain G362C.

[1870] A variant glucosidase may contain T365A, T365N or T365S, in particular T365N.

[1871] A variant glucosidase may contain L367C.

[1872] A variant glucosidase may contain V394R.

[1873] A variant glucosidase may contain V395Y.

[1874] A variant glucosidase may contain Q396E, Q396G, Q396N, Q396P, Q396R, Q396S or Q396Y, in particular Q396R.

[1875] A variant glucosidase may contain F430W.

[1876] A variant glucosidase may contain R435F.

[1877] A variant glucosidase may contain V438T.

[1878] A variant glucosidase may contain V440F.

[1879] A variant glucosidase may contain F442M or F442Q, in particular F442Q.

[1880] A variant glucosidase may contain G443D.

[1881] A variant glucosidase may contain G444T.

[1882] A variant glucosidase may contain A473F or A473R, in particular A473F.

[1883] A variant glucosidase may contain L474C, L474I or L474V, in particular L474C.

[1884] A variant glucosidase may contain I475F.

[1885] A variant glucosidase may contain L492C, L492G, L492H, L492I, L492N, L492Q, L492V, L492W or L492Y, in particular L492H, L492N, L492V. In some embodiments a variant glucosidase contains L492H. In some embodiments, a variant glucosidase contains L492N. In some embodiments a variant glucosidase contains L492V.

[1886] A variant glucosidase may contain Q493F or Q493H.

[1887] A variant glucosidase may contain P494H or P494I, in particular P494I.

[1888] A variant glucosidase may contain S495I, S495K or S495Q.

[1889] A variant glucosidase may contain G496P or G496W, in particular G496P.

[1890] A variant glucosidase may contain D498A, D498E, D498F, D498I, D498K, D498L, D498N, D498P, D498R, D498S, D498T or D498V, in particular D498P.

[1891] A variant glucosidase may contain A502R.

[1892] A variant glucosidase may contain M504G or M504R, in particular M504R.

[1893] A variant glucosidase may contain L507A or L507R, in particular L507R.

[1894] A variant glucosidase may contain T508M.

[1895] A variant glucosidase may contain L529M.

[1896] A variant glucosidase may contain F535P.

[1897] A variant glucosidase may contain A536D or A536E.

[1898] A variant glucosidase may contain A537R.

[1899] A variant glucosidase may contain F541A, F541I, F541L, F541M or F541V, in particular F541I.

[1900] A variant glucosidase may contain L542I.

[1901] A variant glucosidase may contain Q543G or Q543L.

[1902] A variant glucosidase may contain E547L.

[1903] A variant glucosidase may contain Y585W.

[1904] A variant glucosidase may contain E588K.

[1905] Variant glucosidases may comprise R357M, T365N, A473F, L474C and I475F.

[1906] Variant glucosidases may comprise F44Y, R357M, T365N, F442Q, A473F, L474C and I475F.

[1907] Variant glucosidases may comprise F44Y, V263L, R357M, T365N, F442Q, A473F, L474C, I475F and F541I.

[1908] Variant glucosidases may comprise F44Y, V263L, A355W, R357M, T365N, L367C, Q396R, F442Q, L474C, I475F and F541I.

[1909] Variant glucosidases may comprise F44Y, V263L, R357M, T365N, F442Q, L474C, I475F, F541I and zero to seventeen mutations selected from the list consisting of: (iii) N351H

[1910] (iv) A355H, A355I, A355L, A355M, A355R, A355T or A355W

[1911] (v) A356P

[1912] (viii) L367C

[1913] (x) G443D

[1914] (xi) A473F

[1915] (xiv) L492H, L492N, L492V

[1916] (xv) P494I

[1917] (xvi) G496P

[1918] (xvii) D498P

[1919] (xviii) M504R

[1920] (xix) L507R

[1921] (xx) F535P

[1922] (xxi) A537R

[1923] (xxiii) L542I

[1924] (xxiv) E547L and

[1925] (xxv) E588K.

[1926] A variant glucosidase may comprise a “tag,” a sequence of amino acids that allows for the isolation and / or identification of the polypeptide. For example, adding an affinity tag can be useful in purification. Exemplary affinity tags that can be used include histidine (HIS) tags (e.g. hexa histidine-tag, or 6×His-Tag), FLAG-TAG, and HA tags. Tags may be located N-terminally or C-terminally and may be directly connected or attached via a linking sequence. SEQ ID No. 1177 provides a sequence for an exemplary 6×His-Tag with linker sequence which may be N-terminally attached. SEQ ID No. 1178 provides a sequence for an exemplary 6×His-Tag with linker sequence which may be C-terminally attached.

[1927] In certain embodiments, the tags used herein are removable, e.g., removal by chemical agents or by enzymatic means, once they are no longer needed, e.g., after the polypeptide has been purified.

[1928] A variant glucosidase may comprise 1000 residues or fewer, especially 950 residues or fewer, in particular 900 residues or fewer, such as 850 residues or fewer.

[1929] A variant glucosidase may consist of an amino acid sequence of SEQ ID No. 262 with one to twenty-five mutations selected from the list consisting of:

[1930] (i) F44Y

[1931] (ii) V263L

[1932] (iii) N351H

[1933] (iv) A355H, A355I, A355L, A355M, A355R, A355T or A355W

[1934] (v) A356P

[1935] (vi) R357M

[1936] (vii) T365N

[1937] (viii) L367C

[1938] (ix) F442Q

[1939] (x) G443D

[1940] (xi) A473F

[1941] (xii) L474C

[1942] (xiii) I475F

[1943] (xiv) L492H, L492N, L492V

[1944] (xv) P494I

[1945] (xvi) G496P

[1946] (xvii) D498P

[1947] (xviii) M504R

[1948] (xix) L507R

[1949] (xx) F535P

[1950] (xxi) A537R

[1951] (xxii) F541I

[1952] (xxiii) L542I

[1953] (xxiv) E547L and

[1954] (xxv) E588K.

[1955] Variant glucosidases desirably demonstrate a FIOP (Fold Improvement Over Parent) relative to SEQ ID No. 262 of at least 1.05, especially at least 2, in particular at least 10, such as at least 50. FIOP may be determined by the methods described in Example 4.

[1956] Kribbella flavida rhamnosidase (Uniparc reference UPI00019BDB13, Uniprot reference D2PMT5—SEQ ID No. 1017 herein) is a naturally occurring rhamnosidase demonstrating alpha exo rhamnosidase activity and, for example, is capable of the conversion of desglucosyl-QS-17 family components to QS-21 family components. Despite its potent activity, the present inventors have found that the properties of wild type Kribbella flavida rhamnosidase may be altered by the introduction of one or more mutations.

[1957] The present application describes an engineered rhamnosidase polypeptide for use in the method of the invention comprising, such as consisting of, an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID No. 1017, or a functional fragment thereof, wherein the engineered rhamnosidase polypeptide includes at least one residue substitution from:

[1958] (i) A56C

[1959] (ii) A143P

[1960] (iii) Q181H, Q181R or Q181S

[1961] (iv) L214M

[1962] (v) G215S

[1963] (vi) F216M

[1964] (vii) G218D or G218N

[1965] (viii) K219G

[1966] (ix) A238M

[1967] (x) T252Y

[1968] (xi) T311W

[1969] (xii) V326C

[1970] (xiii) G357C

[1971] (xiv) S369C, S369I, S369K or S369M

[1972] (xv) I487M, I487Q or I487V

[1973] (xvi) K492N

[1974] (xvii) V499T

[1975] (xviii) G508S

[1976] (xix) R543C

[1977] (xx) L557Y

[1978] (xxi) G634A

[1979] (xxii) S635N

[1980] (xxiii) A690C and

[1981] (xxiv) Q921H.

[1982] Consequently, the present invention provides a polypeptide comprising an amino acid sequence of sequence of SEQ ID No. 1017 with one to twenty-four mutations selected from the list consisting of:

[1983] (i) A56C

[1984] (ii) A143P

[1985] (iii) Q181H, Q181R or Q181S

[1986] (iv) L214M

[1987] (v) G215S

[1988] (vi) F216M

[1989] (vii) G218D or G218N

[1990] (viii) K219G

[1991] (ix) A238M

[1992] (x) T252Y

[1993] (xi) T311W

[1994] (xii) V326C

[1995] (xiii) G357C

[1996] (xiv) S369C, S369I, S369K or S369M

[1997] (xv) I487M, I487Q or I487V

[1998] (xvi) K492N

[1999] (xvii) V499T

[2000] (xviii) G508S

[2001] (xix) R543C

[2002] (xx) L557Y

[2003] (xxi) G634A

[2004] (xxii) S635N

[2005] (xxiii) A690C and

[2006] (xxiv) Q921H.

[2007] Such polypeptides may be referred to herein as ‘variant rhamnosidases’.

[2008] Variant rhamnosidases will contain one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three or all twenty-four mutations.

[2009] A variant rhamnosidase may contain A56C.

[2010] A variant rhamnosidase may contain A143P.

[2011] A variant rhamnosidase may contain Q181H, Q181R or Q181S. In some embodiments a variant rhamnosidase contains Q181H. In some embodiments a variant rhamnosidase contains Q181R. In some embodiments a variant rhamnosidase contains Q181S.

[2012] A variant rhamnosidase may contain L214M.

[2013] A variant rhamnosidase may contain G215S.

[2014] A variant rhamnosidase may contain F216M.

[2015] A variant rhamnosidase may contain G218D or G218N. In some embodiments a variant rhamnosidase contains G218D. In some embodiments a variant rhamnosidase contains G218N.

[2016] A variant rhamnosidase may contain K219G.

[2017] A variant rhamnosidase may contain A238M.

[2018] A variant rhamnosidase may contain T252Y.

[2019] A variant rhamnosidase may contain T311W.

[2020] A variant rhamnosidase may contain V326C.

[2021] A var...

Claims

1. A method for making a product saponin, the method comprising:(i) providing a plant cell culture extract comprising saponins; and(ii) enzymatically converting a starting saponin from the plant cell culture extract to the product saponin.

2. A method for making a product saponin, the method comprising:(i) culturing plant cells that are capable of synthesis of saponins under conditions leading to the synthesis of the saponins;(ii) recovering the saponins from the plant cell culture; and(iii) enzymatically converting a starting saponin from the recovered saponins to the product saponin.

3. A method for making a product saponin, the method comprising:(i) culturing plant cells that are capable of synthesis of saponins under conditions leading to the synthesis of the saponins;(ii) enzymatically converting a starting saponin from the synthesized saponins to the product saponin; and(iii) recovering saponins from the plant cell culture.

4. A method for increasing an amount of a product saponin obtainable from a plant cell culture, the method comprising:(i) culturing plant cells that are capable of synthesis of saponins under conditions leading to the synthesis of the saponins;(ii) recovering the saponins from the plant cell culture; and(iii) enzymatically converting a starting saponin from the recovered saponins to the product saponin.

5. A method for increasing an amount of a product saponin obtainable from a plant cell culture, the method comprising:(i) culturing plant cells that are capable of synthesis of saponins under conditions leading to the synthesis of the saponins;(ii) enzymatically converting a starting saponin from the synthesized saponins to the product saponin; and(iii) recovering saponins from the plant cell culture.

6. (canceled)7. (canceled)8. A method for producing saponins by plant cell culture, the method comprising:(i) culturing plant cells that are capable of synthesis of the saponins under conditions leading to the synthesis of the saponins; and(ii) recovering saponins,wherein the yield of a product saponin is increased by enzymatically converting a starting saponin from the recovered saponins to the product saponin.

9. The method of claim 1, wherein the saponins are quillaic acid glycosides.

10. The method of claim 1 wherein the plant cell culture extract are from Quillaja saponaria.

11. The method of claim 10, wherein the starting saponin is a QS-18 family component.

12. The method of claim 10, wherein the starting saponin is a desglucosyl-QS-17 family component.

13. The method of claim 10, wherein the starting saponin is a QS-17 family component.

14. The method of claim 10, wherein the starting saponin is a desarabinofuranosyl-QS-18 family component.

15. The method of claim 10, wherein the starting saponin is an acetylated desglucosyl-QS-17 family component.

16. The method of claim 11, wherein the product saponin is a QS-21 family component.

17. The method of claim 13, wherein the product saponin is a QS-18 family component or a desglucosyl-QS-17 family component.

18. (canceled)19. The method of claim 14, wherein the product saponin is a desarabinofuranosyl-QS-21 family component.

20. The method of claim 15, wherein the product saponin is an acetylated QS-21 family component.

21. The method of claim 1, wherein one single starting saponin is converted to one single product saponin.

22. The method of claim 1, wherein a plurality of starting saponins is converted to a plurality of product saponins.

23. The method of claim 1, wherein the enzymatic conversion comprises the removal of a beta-glucose residue on the starting saponin by a glucosidase.

24. The method of claim 23, wherein the glucosidase comprises SEQ ID NO: 262, SEQ ID NO: 208, SEQ ID NO: 63, SEQ ID NO: 229, SEQ ID NO: 250, SEQ ID NO: 5, SEQ ID NO: 101, SEQ ID NO: 207, SEQ ID NO: 169, SEQ ID NO: 247, SEQ ID NO: 302, SEQ ID NO: 324, SEQ ID NO: 319, SEQ ID NO: 9, SEQ ID NO: 240, SEQ ID NO: 325, SEQ ID NO: 338, SEQ ID NO: 850, SEQ ID NO: 879, SEQ ID NO: 868, SEQ ID NO: 826, SEQ ID NO: 804, SEQ ID NO: 888, SEQ ID NO: 881, SEQ ID NO: 891, SEQ ID NO: 816, SEQ ID NO: 827, SEQ ID NO: 857, SEQ ID NO: 853, SEQ ID NO: 842, SEQ ID NO: 814, SEQ ID NO: 886, SEQ ID NO: 885, SEQ ID NO: 838, SEQ ID NO: 829, SEQ ID NO: 808, SEQ ID NO: 828, SEQ ID NO: 870, SEQ ID NO: 873, SEQ ID NO: 844, SEQ ID NO: 882, SEQ ID NO: 874, SEQ ID NO: 825, SEQ ID NO: 824, SEQ ID NO: 823, SEQ ID NO: 810, SEQ ID NO: 894, SEQ ID NO: 849, SEQ ID NO: 803, SEQ ID NO: 890, SEQ ID NO: 841, SEQ ID NO: 832, SEQ ID NO: 830, SEQ ID NO: 845, SEQ ID NO: 871, SEQ ID NO: 837, SEQ ID NO: 883, SEQ ID NO: 809, or a functional variant thereof.

25. The method of claim 1, wherein the enzymatic conversion involves the removal of an alpha-rhamnose residue on the starting saponin by a rhamnosidase.

26. The method of claim 25, wherein the rhamnosidase comprises SEQ ID NO: 992, SEQ ID NO: 1003, SEQ ID NO: 1052, SEQ ID NO: 1073, SEQ ID NO: 1017, SEQ ID NO: 1055, SEQ ID NO: 1075, SEQ ID NO: 1001, SEQ ID NO: 1007, SEQ ID NO: 1061, SEQ ID NO: 1079, SEQ ID NO: 1027, SEQ ID NO: 1039, SEQ ID NO: 1041, SEQ ID NO: 989, SEQ ID NO: 1053, SEQ ID NO: 1018, SEQ ID NO: 1066, SEQ ID NO: 1082, SEQ ID NO: 1076, SEQ ID NO: 993, SEQ ID NO: 1077, SEQ ID NO: 1046, SEQ ID NO: 1015, SEQ ID NO: 1063, SEQ ID NO: 1054, SEQ ID NO: 1074, SEQ ID NO: 1067, SEQ ID NO: 1033, or a functional variant thereof.

27. (canceled)28. (canceled)29. (canceled)30. (canceled)31. (canceled)

Images